This document provides an introduction to animal cell culture. It discusses what cell and tissue culture are, how cell cultures are obtained through primary culture or purchasing established cell lines. It describes the characteristics of cultured cells, including their morphology and functional properties. Some of the key challenges in cell culture are avoiding contamination and providing an optimal environment for cell growth. The document also gives a brief overview of what cell culture is used for.
Tissue culture involves growing cells, tissues, or organs outside of their natural biological context. There are three main types of tissue culture: cell culture, explant culture, and organ culture. Cell culture involves growing isolated cells, either in an adherent monolayer or in suspension. Eplant culture grows fragments of tissue on a substrate, while organ culture maintains the three-dimensional structure of whole organs. Tissue culture has advantages like control over the environment and scale-up potential, but cells may lose differentiated characteristics over time.
This document discusses cell culture in animal biotechnology. It provides an overview of the history of cell culture from the late 1800s to modern applications. Some key points covered include:
1) Cell culture refers to growing cells in a controlled artificial environment outside of their natural environment. Examples of cell types grown include fibroblasts, lymphocytes, and cells from various tissues.
2) Important milestones in the history of cell culture include Roux maintaining embryonic chick cells in 1885 and Carrel introducing strict aseptic techniques allowing long-term cell culture in 1913.
3) Cell culture has various applications including use as model systems, in toxicity testing, cancer research, virology, drug development, and gene therapy.
This document discusses animal tissue culture. It defines animal tissue culture as the scientific process of growing animal tissue cells outside the body in a nutrient-rich medium. The document outlines the historical background of tissue culture, requirements for culture, types of cells cultured, steps to culture tissue, types of culture (primary, subculture, cell line), common contaminants (bacteria, viruses, yeast), and applications/merits of tissue culture like virology, manufacturing, research, and gene therapy. Limitations include needing expertise to interpret cell behavior and higher costs compared to whole tissue studies.
This document discusses animal tissue culture and cell culture techniques. It begins by defining tissue culture as the removal and growth of cells, tissues or organs from animals or plants in an artificial environment that supplies nutrients for growth. It then covers major developments in the field like the use of antibiotics, trypsin for subculturing, and chemically defined media. Applications of cell culture discussed include research areas like toxicology testing, cancer research, virology and genetic engineering. The document also covers primary culture, cell lines, monolayer and suspension culture systems, culturing adherent and suspension cells, cryopreservation of cells, cell viability assessment, and basic cell culture equipment.
This lecture discusses animal cell biotechnology and cell culture. It begins by outlining some key applications of animal cell cultures, including for viral vaccines, monoclonal antibodies, recombinant proteins, and hormones. It then describes the basic concepts of cell culture, including how cells can continue growing when removed from tissue and supplied with nutrients. The lecture covers important historical figures like Harrison, Carrel, Hayflick and Moorhead and their key contributions to the field. It discusses the finite lifespan of cultured cells and how they can become immortal through transformation. The characteristics of normal versus transformed cells are also summarized.
Primary cell cultures are derived directly from animal tissues and have a limited lifespan, usually undergoing fewer than 10 divisions. They retain characteristics of the original tissue. Diploid cell strains can undergo 20-50 passages while maintaining the original karyotype. Continuous cell lines are immortalized cell lines that can divide indefinitely, having undergone changes including aneuploidy and loss of differentiation. Common types of cell culture include primary cultures from tissues like monkey kidney, diploid strains from fetal tissues like human lung fibroblasts, and continuous lines derived from tumors.
Tissue culture involves growing cells, tissues, or organs outside of their natural biological context. There are three main types of tissue culture: cell culture, explant culture, and organ culture. Cell culture involves growing isolated cells, either in an adherent monolayer or in suspension. Eplant culture grows fragments of tissue on a substrate, while organ culture maintains the three-dimensional structure of whole organs. Tissue culture has advantages like control over the environment and scale-up potential, but cells may lose differentiated characteristics over time.
This document discusses cell culture in animal biotechnology. It provides an overview of the history of cell culture from the late 1800s to modern applications. Some key points covered include:
1) Cell culture refers to growing cells in a controlled artificial environment outside of their natural environment. Examples of cell types grown include fibroblasts, lymphocytes, and cells from various tissues.
2) Important milestones in the history of cell culture include Roux maintaining embryonic chick cells in 1885 and Carrel introducing strict aseptic techniques allowing long-term cell culture in 1913.
3) Cell culture has various applications including use as model systems, in toxicity testing, cancer research, virology, drug development, and gene therapy.
This document discusses animal tissue culture. It defines animal tissue culture as the scientific process of growing animal tissue cells outside the body in a nutrient-rich medium. The document outlines the historical background of tissue culture, requirements for culture, types of cells cultured, steps to culture tissue, types of culture (primary, subculture, cell line), common contaminants (bacteria, viruses, yeast), and applications/merits of tissue culture like virology, manufacturing, research, and gene therapy. Limitations include needing expertise to interpret cell behavior and higher costs compared to whole tissue studies.
This document discusses animal tissue culture and cell culture techniques. It begins by defining tissue culture as the removal and growth of cells, tissues or organs from animals or plants in an artificial environment that supplies nutrients for growth. It then covers major developments in the field like the use of antibiotics, trypsin for subculturing, and chemically defined media. Applications of cell culture discussed include research areas like toxicology testing, cancer research, virology and genetic engineering. The document also covers primary culture, cell lines, monolayer and suspension culture systems, culturing adherent and suspension cells, cryopreservation of cells, cell viability assessment, and basic cell culture equipment.
This lecture discusses animal cell biotechnology and cell culture. It begins by outlining some key applications of animal cell cultures, including for viral vaccines, monoclonal antibodies, recombinant proteins, and hormones. It then describes the basic concepts of cell culture, including how cells can continue growing when removed from tissue and supplied with nutrients. The lecture covers important historical figures like Harrison, Carrel, Hayflick and Moorhead and their key contributions to the field. It discusses the finite lifespan of cultured cells and how they can become immortal through transformation. The characteristics of normal versus transformed cells are also summarized.
Primary cell cultures are derived directly from animal tissues and have a limited lifespan, usually undergoing fewer than 10 divisions. They retain characteristics of the original tissue. Diploid cell strains can undergo 20-50 passages while maintaining the original karyotype. Continuous cell lines are immortalized cell lines that can divide indefinitely, having undergone changes including aneuploidy and loss of differentiation. Common types of cell culture include primary cultures from tissues like monkey kidney, diploid strains from fetal tissues like human lung fibroblasts, and continuous lines derived from tumors.
This document discusses basic concepts of animal cell culture. It begins with an introduction and brief history of cell culture. Some key advancements include the use of antibiotics to reduce contamination, trypsin to subculture adherent cells, and chemically defined culture media. Current applications of cell culture include cancer research, genetic engineering, and gene therapy. The document then covers various cell culture types (primary vs cell line), requirements, techniques, and factors affecting growth. It distinguishes between adherent and suspension cultures as well as finite and continuous cell lines.
The document discusses different types of cell culture used in bioreactors. It describes organ culture, tissue culture, and cell culture. Cell culture involves dispersing tissue enzymatically into a cell suspension that can be grown as a monolayer or in suspension. Continuous cell lines can be propagated indefinitely and have gained immortality through transformation. Bioreactors must provide a well-controlled environment for cell culture and can operate in batch, fed-batch or perfusion modes. Common bioreactor designs include stirred tank, airlift and wave bioreactors.
This document discusses animal cell culture techniques. It describes primary cell culture which uses cells directly from tissue and is heterogeneous and finite. Secondary culture is produced by sub-culturing primary cells, which can become cell lines that are homogeneous and can divide indefinitely. Cell lines are categorized as finite, which senesce after limited divisions, or continuous, which can divide indefinitely. Common cell lines discussed include MCF-7, HL-60, and HeLa. Requirements for cell culture include media and lab equipment, and it aims to maintain pH between 7.2-7.4.
Animal cell culture media typically contain energy sources like glucose, amino acids as nitrogen sources, vitamins, inorganic salts, fatty acids, antibiotics, growth factors, and hormones. Most media also require an incubator to maintain optimal temperature, pH, osmolality, and gaseous environment for cell growth. Cell cultures can be grown adhered to surfaces or in suspension, and may have limited or continuous proliferation. Common applications of animal cell culture include vaccine production, cancer research, pharmaceutical drug production, and studying nerve cell function.
Cultured animal cells have many important applications. They can be used as (1) model systems to study basic cell biology and interactions between cells and pathogens, (2) for toxicity testing of new drugs and chemicals, and (3) in cancer research to study normal and cancerous cell differences. Animal cell culture is also used for virology research, manufacturing of vaccines and proteins, genetic counseling, genetic engineering of cells, and gene and drug screening and development. Proper growth media, aseptic techniques, cryopreservation, and applications in various fields make animal cell culture a valuable tool.
Cell lines are cultured by removing cells from tissue and growing them in a favorable artificial environment. Primary cell cultures refer to the initial proliferation of isolated cells on a substrate. When the primary culture is subcultured to a new vessel, it becomes a cell line. Cell lines are either finite, with limited growth ability before senescence, or continuous, having undergone transformation allowing indefinite division. Cell strains are subpopulations of cell lines selected through cloning or other methods.
Mammalian cell culture was first developed in the early 20th century and has since enabled significant advances in research areas like drug development. Proper aseptic technique and regular maintenance are required to prevent contamination and allow cells to grow healthily. Contamination can occur chemically from unwanted substances or microbially from bacteria, fungi, viruses, and mycoplasma. Strict adherence to safety policies and prevention of cross-contamination between cell lines are important to ensure sterile conditions and validate experimental results.
Introduction
Primary Culture
Steps In Primary Culture
Isolation Of Tissue
Dissection And/Or Disaggregation
Types Of Primary Culture
Primary Explant Culture
Enzymatic Disaggregation
Mechanical Disaggregation
Cell Line( Finite & Continuous)
Naming A Cell Line
Choosing A Cell Line
Maintenance Of Cell Line
Conclusion
reference
This presentation covers the introduction to Insect Cell Culture. Also covers its general information about cell culture practices followed in the lab. It covers culture media, the source of cells for culture and examples of the cell line with their culture conditions.
For decades, cell lines have played a critical role in scientific developments. In most cases, researchers just got data generated from cell lines. However, due to some weaknesses of cell lines, scientists become increasingly cautious about these generated results. But now the game has changed! Primary cells now are believed to be a more biologically relevant tool than cell lines for studying human and animal biology. And we design this primary cell culture guide aimed at showing new investigators the basic principles of primary cell and some practical culture skills.
Cell culture is the process of growing cells outside their natural environment. Key events in the history of cell culture include Wilhelm Roux maintaining chicken tissue in saline in 1885, and Harrison establishing the first cell culture in 1907. Traditionally, cells were cultured in 2D monolayers, but 3D cell culture has emerged as a way to better mimic the in vivo microenvironment. 3D cultures can be scaffold-based, using matrices like collagen, or scaffold-free, allowing cell-cell interactions. Technologies like microfluidics and bio-MEMS now aid 3D cell culture research. Careful planning is required to properly design and safely operate a cell culture laboratory.
History of animal tissue culture and natural surroundings for animal cellNeeraj Chauhan
This document discusses the history of animal tissue culture and factors that affect culturing animal cells. It notes that Roux in 1885 was the first to culture embryonic chick cells and Harrison in 1907 successfully cultured nerve cells. Key events over 130 years included development of defined media and serum-free media. Factors that impact cell culture choice include cell yield, whether cells are monolayer or suspension, venting, sampling needs, growth uniformity, and cost. Environmental factors like pH, temperature, gas phase, osmolarity, foaming, and viscosity must also be controlled to maintain optimal cell growth conditions.
Cell culture involves growing cells from tissue or organ samples in artificial environments outside of the original organism. There are several stages of cell culture, beginning with isolating tissues through enzymatic or mechanical means. Primary cell cultures have a limited lifespan, while continuous cell lines can proliferate indefinitely. Proper culture conditions require appropriate media, substrates, gases, and temperature/humidity control. Cells may be grown as adherent monolayers or in suspension. Cell culture has many applications including drug development, cancer research, and production of therapeutic products.
Javier Amayra - Biotechnological Screening in Animal Cell Cultureponenciasexpoquim11
The document discusses bioprocess development for animal cell culture. It highlights that screening phases are important but have reproducibility issues when scaling up. The HexaScreen and HexaBatch systems aim to address these issues by providing (1) an automated and controlled multi-vessel screening platform, (2) low volume bioreactors to better mimic industrial scales, and (3) monitoring of key parameters like cell growth, pH and oxygen to improve screening accuracy and translation to later phases. This helps streamline bioprocess optimization from initial screening through production.
1. The history of animal cell culture began in 1907 with Harrison cultivating frog nerve cells. Over the decades, techniques improved including the development of defined media, antibiotics in culture, and the first human cell line in 1952.
2. Cell culture media contains essential nutrients, growth factors, hormones, and other components to support cell growth. Basal media can be supplemented with serum, serum-free, or reduced serum. Common basal media include DMEM and RPMI.
3. There are three main types of tissue culture: explant culture uses intact pieces of tissue; organ culture maintains tissue structure; and cell culture dissociates cells from tissue into single cell suspensions. Each technique offers advantages and disadvantages depending on
This document provides an overview of animal cell and tissue culture. It begins by defining animal cell culture as dealing with controlling and modulating cellular function, recognizing cells as independent organisms. Examples are given of transforming cells into engineered systems to express specific biological functions in vitro. Considerations for animal cell culture models include choosing appropriate cell types and designing systems that mimic human physiology. Requirements for setting up an animal cell culture like media, incubators, and safety equipment are also outlined. Specific techniques are discussed such as obtaining skin cells and culturing types of liver cells. Requirements for the next class on differences between animal and cell experiments are assigned.
This document provides an introduction to animal cell culture by Dr. Anu P. Abhimannue. It discusses the history and development of animal cell culture from the early 20th century. It describes different types of animal cell culture such as primary versus secondary culture and finite versus continuous cell lines. It also discusses various cell culture methods like monolayer, suspension, types of culture vessels used and morphology of cultured cells. The document provides advantages and limitations of animal cell culture techniques.
This document discusses different types of mammalian cell culture. It describes primary cell culture, which uses cells directly from tissue that can undergo a limited number of divisions before senescing. Finite and continuous cell lines can proliferate for extended periods through transformation or immortalization. Common cell lines include HeLa cells and other tumor-derived lines. The document also covers techniques for attachment and suspension cell culture, and factors that influence cell growth in vitro.
This chapter discusses cell culture techniques for isolating animal viruses. It defines key terms like cell line, strain, cloning and outlines best practices. Primary importance is given to obtaining low passage cells from reputable suppliers with documentation. The chapter notes that while technology has advanced, emphasis on cell culture as an art is lacking. Proper training and standardized practices are needed for reliable, repeatable results.
Cell and tissue culture involves removing cells or tissues from living organisms and placing them in an artificial environment conducive to growth. This environment typically consists of a glass or plastic vessel containing a liquid or semisolid medium supplying necessary nutrients. There are two main methods for obtaining cell cultures - explant culture, which involves attaching tissue fragments to a culture vessel, and enzymatic dissociation, which uses enzymes like trypsin to separate cells. Maintaining cell cultures requires specialized equipment like incubators, laminar flow hoods, and microscopes, as well as sterile culture procedures and defined media tailored to cell needs. Cell and tissue cultures have many applications, including cancer research, virology, genetic counseling, and gene therapy.
This document discusses basic concepts of animal cell culture. It begins with an introduction and brief history of cell culture. Some key advancements include the use of antibiotics to reduce contamination, trypsin to subculture adherent cells, and chemically defined culture media. Current applications of cell culture include cancer research, genetic engineering, and gene therapy. The document then covers various cell culture types (primary vs cell line), requirements, techniques, and factors affecting growth. It distinguishes between adherent and suspension cultures as well as finite and continuous cell lines.
The document discusses different types of cell culture used in bioreactors. It describes organ culture, tissue culture, and cell culture. Cell culture involves dispersing tissue enzymatically into a cell suspension that can be grown as a monolayer or in suspension. Continuous cell lines can be propagated indefinitely and have gained immortality through transformation. Bioreactors must provide a well-controlled environment for cell culture and can operate in batch, fed-batch or perfusion modes. Common bioreactor designs include stirred tank, airlift and wave bioreactors.
This document discusses animal cell culture techniques. It describes primary cell culture which uses cells directly from tissue and is heterogeneous and finite. Secondary culture is produced by sub-culturing primary cells, which can become cell lines that are homogeneous and can divide indefinitely. Cell lines are categorized as finite, which senesce after limited divisions, or continuous, which can divide indefinitely. Common cell lines discussed include MCF-7, HL-60, and HeLa. Requirements for cell culture include media and lab equipment, and it aims to maintain pH between 7.2-7.4.
Animal cell culture media typically contain energy sources like glucose, amino acids as nitrogen sources, vitamins, inorganic salts, fatty acids, antibiotics, growth factors, and hormones. Most media also require an incubator to maintain optimal temperature, pH, osmolality, and gaseous environment for cell growth. Cell cultures can be grown adhered to surfaces or in suspension, and may have limited or continuous proliferation. Common applications of animal cell culture include vaccine production, cancer research, pharmaceutical drug production, and studying nerve cell function.
Cultured animal cells have many important applications. They can be used as (1) model systems to study basic cell biology and interactions between cells and pathogens, (2) for toxicity testing of new drugs and chemicals, and (3) in cancer research to study normal and cancerous cell differences. Animal cell culture is also used for virology research, manufacturing of vaccines and proteins, genetic counseling, genetic engineering of cells, and gene and drug screening and development. Proper growth media, aseptic techniques, cryopreservation, and applications in various fields make animal cell culture a valuable tool.
Cell lines are cultured by removing cells from tissue and growing them in a favorable artificial environment. Primary cell cultures refer to the initial proliferation of isolated cells on a substrate. When the primary culture is subcultured to a new vessel, it becomes a cell line. Cell lines are either finite, with limited growth ability before senescence, or continuous, having undergone transformation allowing indefinite division. Cell strains are subpopulations of cell lines selected through cloning or other methods.
Mammalian cell culture was first developed in the early 20th century and has since enabled significant advances in research areas like drug development. Proper aseptic technique and regular maintenance are required to prevent contamination and allow cells to grow healthily. Contamination can occur chemically from unwanted substances or microbially from bacteria, fungi, viruses, and mycoplasma. Strict adherence to safety policies and prevention of cross-contamination between cell lines are important to ensure sterile conditions and validate experimental results.
Introduction
Primary Culture
Steps In Primary Culture
Isolation Of Tissue
Dissection And/Or Disaggregation
Types Of Primary Culture
Primary Explant Culture
Enzymatic Disaggregation
Mechanical Disaggregation
Cell Line( Finite & Continuous)
Naming A Cell Line
Choosing A Cell Line
Maintenance Of Cell Line
Conclusion
reference
This presentation covers the introduction to Insect Cell Culture. Also covers its general information about cell culture practices followed in the lab. It covers culture media, the source of cells for culture and examples of the cell line with their culture conditions.
For decades, cell lines have played a critical role in scientific developments. In most cases, researchers just got data generated from cell lines. However, due to some weaknesses of cell lines, scientists become increasingly cautious about these generated results. But now the game has changed! Primary cells now are believed to be a more biologically relevant tool than cell lines for studying human and animal biology. And we design this primary cell culture guide aimed at showing new investigators the basic principles of primary cell and some practical culture skills.
Cell culture is the process of growing cells outside their natural environment. Key events in the history of cell culture include Wilhelm Roux maintaining chicken tissue in saline in 1885, and Harrison establishing the first cell culture in 1907. Traditionally, cells were cultured in 2D monolayers, but 3D cell culture has emerged as a way to better mimic the in vivo microenvironment. 3D cultures can be scaffold-based, using matrices like collagen, or scaffold-free, allowing cell-cell interactions. Technologies like microfluidics and bio-MEMS now aid 3D cell culture research. Careful planning is required to properly design and safely operate a cell culture laboratory.
History of animal tissue culture and natural surroundings for animal cellNeeraj Chauhan
This document discusses the history of animal tissue culture and factors that affect culturing animal cells. It notes that Roux in 1885 was the first to culture embryonic chick cells and Harrison in 1907 successfully cultured nerve cells. Key events over 130 years included development of defined media and serum-free media. Factors that impact cell culture choice include cell yield, whether cells are monolayer or suspension, venting, sampling needs, growth uniformity, and cost. Environmental factors like pH, temperature, gas phase, osmolarity, foaming, and viscosity must also be controlled to maintain optimal cell growth conditions.
Cell culture involves growing cells from tissue or organ samples in artificial environments outside of the original organism. There are several stages of cell culture, beginning with isolating tissues through enzymatic or mechanical means. Primary cell cultures have a limited lifespan, while continuous cell lines can proliferate indefinitely. Proper culture conditions require appropriate media, substrates, gases, and temperature/humidity control. Cells may be grown as adherent monolayers or in suspension. Cell culture has many applications including drug development, cancer research, and production of therapeutic products.
Javier Amayra - Biotechnological Screening in Animal Cell Cultureponenciasexpoquim11
The document discusses bioprocess development for animal cell culture. It highlights that screening phases are important but have reproducibility issues when scaling up. The HexaScreen and HexaBatch systems aim to address these issues by providing (1) an automated and controlled multi-vessel screening platform, (2) low volume bioreactors to better mimic industrial scales, and (3) monitoring of key parameters like cell growth, pH and oxygen to improve screening accuracy and translation to later phases. This helps streamline bioprocess optimization from initial screening through production.
1. The history of animal cell culture began in 1907 with Harrison cultivating frog nerve cells. Over the decades, techniques improved including the development of defined media, antibiotics in culture, and the first human cell line in 1952.
2. Cell culture media contains essential nutrients, growth factors, hormones, and other components to support cell growth. Basal media can be supplemented with serum, serum-free, or reduced serum. Common basal media include DMEM and RPMI.
3. There are three main types of tissue culture: explant culture uses intact pieces of tissue; organ culture maintains tissue structure; and cell culture dissociates cells from tissue into single cell suspensions. Each technique offers advantages and disadvantages depending on
This document provides an overview of animal cell and tissue culture. It begins by defining animal cell culture as dealing with controlling and modulating cellular function, recognizing cells as independent organisms. Examples are given of transforming cells into engineered systems to express specific biological functions in vitro. Considerations for animal cell culture models include choosing appropriate cell types and designing systems that mimic human physiology. Requirements for setting up an animal cell culture like media, incubators, and safety equipment are also outlined. Specific techniques are discussed such as obtaining skin cells and culturing types of liver cells. Requirements for the next class on differences between animal and cell experiments are assigned.
This document provides an introduction to animal cell culture by Dr. Anu P. Abhimannue. It discusses the history and development of animal cell culture from the early 20th century. It describes different types of animal cell culture such as primary versus secondary culture and finite versus continuous cell lines. It also discusses various cell culture methods like monolayer, suspension, types of culture vessels used and morphology of cultured cells. The document provides advantages and limitations of animal cell culture techniques.
This document discusses different types of mammalian cell culture. It describes primary cell culture, which uses cells directly from tissue that can undergo a limited number of divisions before senescing. Finite and continuous cell lines can proliferate for extended periods through transformation or immortalization. Common cell lines include HeLa cells and other tumor-derived lines. The document also covers techniques for attachment and suspension cell culture, and factors that influence cell growth in vitro.
This chapter discusses cell culture techniques for isolating animal viruses. It defines key terms like cell line, strain, cloning and outlines best practices. Primary importance is given to obtaining low passage cells from reputable suppliers with documentation. The chapter notes that while technology has advanced, emphasis on cell culture as an art is lacking. Proper training and standardized practices are needed for reliable, repeatable results.
Cell and tissue culture involves removing cells or tissues from living organisms and placing them in an artificial environment conducive to growth. This environment typically consists of a glass or plastic vessel containing a liquid or semisolid medium supplying necessary nutrients. There are two main methods for obtaining cell cultures - explant culture, which involves attaching tissue fragments to a culture vessel, and enzymatic dissociation, which uses enzymes like trypsin to separate cells. Maintaining cell cultures requires specialized equipment like incubators, laminar flow hoods, and microscopes, as well as sterile culture procedures and defined media tailored to cell needs. Cell and tissue cultures have many applications, including cancer research, virology, genetic counseling, and gene therapy.
This document provides instructions for five plant tissue culture experiments. Experiment 1 demonstrates the totipotency and nutritional requirements of shoot tip and root tip explants from aseptically germinated seedlings. When transferred to different media, the explants show varying growth responses correlated to the media contents. Experiment 2 examines the effects of hormone balance on explant growth and morphogenesis. Experiments 3-5 involve callus formation, suspension cultures, and anther culture techniques. The document provides detailed background information and step-by-step methods for each experiment.
Essay on Plant Tissue Culture Contents:
the Definition of Plant Tissue Culture.
the History of Plant Tissue Culture.
the Basic Requirements of Plant Tissue Culture.
the General Techniques of Plant Tissue Culture.
the Basic Aspects of Plant Tissue Culture.
the Cellular Totipotency.
the Differentiation.
the Methods in Plant Tissue Culture.
the Applications of Plant Tissue Culture.
the Morphogenesis.
the Subculture or Secondary Cell Culture.
the Soma-Clonal Variation.
the Somatic Hybrids and Cybrids.
the Micro-Propagation.
the Artificial Seed.
the Cryopreservation.
Plant bio 1 introduction to cell tissue cultureDr. Preeti Pal
Tissue culture is a method of growing plant cells, tissues or organs in vitro on artificial nutrient media under sterile conditions. Plant tissue culture involves exposing plant tissue to specific nutrients, hormones and light to produce many new cloned plants over a short period. The father of plant tissue culture is considered to be German botanist Gottlieb Haberlandt who conceived the concept of cell culture in 1902. A key aspect of plant tissue culture is initiation and maintenance of callus cultures, which are masses of unorganized proliferating cells grown on artificial media.
To achieve the target of creating a new plant or a plant with desired characteristics, tissue culture is often coupled with recombinant DNA technology. The techniques of plant tissue culture have largely helped in the green revolution by improving the crop yield and quality.
The knowledge obtained from plant tissue cultures has contributed to our understanding of metabolism, growth, differentiation and morphogenesis of plant cells. Further, developments in tissue culture have helped to produce several pathogen-free plants, besides the synthesis of many biologically important compounds, including pharmaceuticals. Because of the wide range of applications, plant tissue culture attracts the attention of molecular biologists, plant breeders and industrialists.
The document discusses the field of tissue engineering and its potential applications. It describes how tissue engineering could help transplant patients by growing organs and help burn victims avoid disfiguring scars by growing new skin and tissues. The key aspect of tissue engineering is developing biomaterials that can act as scaffolds for cell growth and regeneration of tissues and organs as needed by the body.
Introduction
Definition
History
Principle
Cell sources
What cells can be used?
Scaffolds
Biomaterials
Bioreactor
How tissue engineering is done?
How does tissue engineering differ from cloning?
Tissue engineering of specific structures
Application of tissue engineering
Limitations
Conclusion
References
The document summarizes the past, present, and future of regenerative tissue engineering. It discusses how the field began in the 1950s-60s by combining cell biology with new materials to generate living tissue components. Major advances included the use of stem cells and development of biocompatible scaffolds. The future of the field involves improved biomaterials that mimic natural extracellular matrix, bioprinting of complex tissues, and using various stem cell sources for cell therapy and organ regeneration to treat aging populations. The market for tissue engineering is estimated to grow substantially in coming years.
Cell culture involves growing cells outside of their natural environment under controlled conditions. The document discusses the history and development of cell culture techniques. It explains the process of explant culture where a small piece of tissue is used to initiate a primary culture. The requirements for maintaining cell cultures such as appropriate nutrients, pH, temperature and regular sub-culturing to prevent overgrowth are also summarized. The major cell culture applications in areas like disease research, toxicology testing and genetic engineering are highlighted in brief.
Plant tissue culture is the process of growing plant cells, tissues or organs in an artificial nutrient medium under sterile conditions. It allows plants to be grown in vitro from small meristematic tissues. There are several types of plant tissue culture including callus culture, organ culture and cell suspension culture. The basic technique involves surface sterilization of explant tissue followed by establishment and subculture of the culture on nutrient media. Callus culture specifically produces an undifferentiated mass of plant cells called a callus from explants. Plant tissue culture has many applications including disease elimination, genetic studies, large scale propagation and crop improvement.
Cell culture involves cultivating cells outside the body in an artificial environment that mimics in vivo conditions. Some key developments include the use of antibiotics to prevent contamination, trypsin to detach adherent cells for subculturing, and chemically defined media. Primary cultures have a finite lifespan while continuous cell lines can divide indefinitely. Tissue is enzymatically or mechanically broken up before culturing, and cells require nutrients, oxygen, pH balance, and humidity to grow.
This document provides an overview of cell lines and cell culture. It discusses what cell lines are, how they are established from primary cultures, and how they can become cell strains. It also describes culture conditions, types of cell lines (finite vs continuous), nomenclature, selecting appropriate cell lines based on experimental needs, biosafety levels, cell culture hood and incubator setup. The document is intended as an introduction to cell lines and basic cell culture techniques.
Tissue engineering aims to repair or replace damaged tissues and organs. It involves growing cells on scaffolds in bioreactors that mimic physiological conditions. There are five basic steps: 1) obtaining cells, 2) expanding cells in culture, 3) seeding cells onto scaffolds with growth factors, 4) incubating to form new tissues, and 5) implanting the new tissues. Stem cells have potential for tissue engineering due to their ability to self-renew and differentiate. Current research applies stem cells to treat diseases like cardiovascular issues, diabetes, and injuries. Tissue engineering has created bioartificial organs like windpipes and is researching pancreases, bladders, cartilage, blood vessels, and bone marrow. It provides hope for solving
The document provides step-by-step instructions for primary cell culture and passaging cells. It describes removing tissues from animals or humans, digesting the tissues to isolate individual cells, and culturing the primary cells in vitro. As the primary cells grow to confluence, they are passaged by treating them with trypsin-EDTA to detach the cells, then transferring them to new culture vessels with fresh medium to continue growing and multiplying. The document lists the main reagents, equipment, and experimental materials needed and provides detailed protocols for primary culture establishment and subsequent cell passaging.
This document provides an overview of biotechnology and various applications. It discusses cloning in animals and plants. Reproductive cloning involves transferring the nucleus of an adult cell into an egg with its nucleus removed. Recombinant DNA technology transfers DNA fragments between organisms. Stem cells can replicate and form complex structures, and may help treat medical conditions. The document outlines the cloning of various animal species over time. It also discusses cloning endangered species, human cloning for therapeutic purposes, and the in vitro fertilization process.
Tissue engineering aims to create functional human tissues for repair or replacement of damaged organs. It involves obtaining cells, expanding them in culture, seeding them onto a scaffold to grow new tissue, and implanting the construct. Stem cells offer potential due to their ability to differentiate and self-renew. Research applications include creating artificial organs like livers, pancreases, and bladders. Challenges remain in vascularizing tissues and preventing immune rejection, but tissue engineering offers hope for treating diseases.
The document provides information on cell biology topics including cell structure, transport processes, and cell division. It contains:
1) Descriptions of plant and animal cell structures and comparisons between eukaryotic and prokaryotic cells.
2) Explanations of three transport processes - diffusion, osmosis, and active transport - and examples of each in cells and organisms.
3) An overview of cell division through mitosis and its role in growth and repair of cells.
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Introduction to animal_cell_culture
1. Introduction to Animal Cell Culture
Technical Bulletin
John A. Ryan, Ph.D. Introduction
Corning Incorporated
Cell culture has become one of the major tools used in the
Life Sciences
life sciences today. This guide is designed to serve as a basic
900 Chelmsford St.
introduction to animal cell culture. It is appropriate for lab-
Lowell, MA 01851
oratory workers who are using it for the first time, as well as
for those who interact with cell culture researchers and who
want a better understanding of the key concepts and termi-
nology in this interesting and rapidly growing field.
Table of Contents
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 What is Cell and Tissue Culture?
Tissue Culture is the general term for the removal of cells,
What is Cell and Tissue Culture? . . . . . . . . . . . . . . . . . . 1
tissues, or organs from an animal or plant and their subse-
How are Cell Cultures Obtained? . . . . . . . . . . . . . . . . . . 2 quent placement into an artificial environment conducive
to growth. This environment usually consists of a suitable
What Are Cultured Cells Like? . . . . . . . . . . . . . . . . . . . . 3 glass or plastic culture vessel containing a liquid or semi-
solid medium that supplies the nutrients essential for sur-
What Are Some of the Problems Faced by
vival and growth. The culture of whole organs or intact
Cultured Cells? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
organ fragments with the intent of studying their continued
How to Decide if Cultured Cells Are “Happy”? . . . . . . . 6 function or development is called Organ Culture. When
the cells are removed from the organ fragments prior to,
What is Cell Culture Used For? . . . . . . . . . . . . . . . . . . . 6 or during cultivation, thus disrupting their normal relation-
ships with neighboring cells, it is called Cell Culture.
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Although animal cell culture was first successfully undertak-
en by Ross Harrison in 1907, it was not until the late 1940’s
to early 1950’s that several developments occurred that made
cell culture widely available as a tool for scientists. First,
there was the development of antibiotics that made it easier
to avoid many of the contamination problems that plagued
earlier cell culture attempts. Second was the development of
2. the techniques, such as the use of trypsin to remove cells from culture vessels, necessary to
obtain continuously growing cell lines (such as HeLa cells). Third, using these cell lines,
scientists were able to develop standardized, chemically defined culture media that made
it far easier to grow cells. These three areas combined to allow many more scientists to use
Additional cell culture
cell, tissue and organ culture in their research.
terminology and usage
information can be found During the 1960’s and 1970’s, commercialization of this technology had further impact on
on the Society for In Vitro cell culture that continues to this day. Companies, such as Corning, began to develop and
Biology web site at sell disposable plastic and glass cell culture products, improved filtration products and mate-
www.sivb.org/edu_
terminology.asp.
rials, liquid and powdered tissue culture media, and laminar flow hoods. The overall result
of these and other continuing technological developments has been a widespread increase in
the number of laboratories and industries using cell culture today.
How Are Cell Cultures Obtained?
Primary Culture
When cells are surgically removed from an
organism and placed into a suitable culture
environment, they will attach, divide and Remove
tissue
grow. This is called a Primary Culture.
There are two basic methods for doing this.
First, for Explant Cultures, small pieces of Mince or
tissue are attached to a glass or treated plastic chop
culture vessel and bathed in culture medium.
After a few days, individual cells will move
from the tissue explant out onto the culture
vessel surface or substrate where they will Digest with
begin to divide and grow. The second, more proteolytic
enzymes
widely used method, speeds up this process by
Fixed and stained human adding digesting (proteolytic) enzymes, such
foreskin explants on the sur- as trypsin or collagenase, to the tissue frag-
face of a 150 mm culture dish. ments to dissolve the cement holding the cells Place in
The explants were cultured for together. This creates a suspension of single culture
approximately two weeks. Two
cells that are then placed into culture vessels
of the nine explants (bottom
left and right corners) failed to containing culture medium and allowed to
grow. The remaining explants grow and divide. This method is called Enzymatic Dissociation
show good growth. Each square Enzymatic Dissociation.
is approximately 2 cm across.
Subculturing
When the cells in the primary culture vessel have grown and filled up all of the available
culture substrate, they must be Subcultured to give them room for continued growth. This
is usually done by removing them as gently as possible from the substrate with enzymes.
These are similar to the enzymes used in obtaining the primary culture and are used to break
the protein bonds attaching the cells to the substrate. Some cell lines can be harvested by
gently scraping the cells off the bottom of the culture vessel. Once released, the cell suspen-
sion can then be subdivided and placed into new culture vessels.
Once a surplus of cells is available, they can be treated with suitable cryoprotective agents,
such as dimethylsulfoxide (DMSO) or glycerol, carefully frozen and then stored at cryo-
genic temperatures (below -130°C) until they are needed. The theory and techniques for
Primary culture from the fish cryopreserving cells are covered in the Corning Technical Bulletin: General Guide for
Poeciliopsis lucida. Embryos Cryogenically Storing Animal Cell Cultures (Ref. 9).
were minced and dissociated
with a trypsin solution. These Buying And Borrowing
cells were in culture for about An alternative to establishing cultures by primary culture is to buy established cell cultures
1 week and have formed a
confluent monolayer.
from organizations such as the ATCC (www.atcc.org), or the Coriell Institute for Medical
Research (ccr.coriell.org). These two nonprofit organizations provide high quality cell lines
that are carefully tested to ensure the authenticity of the cells.
2
3. More frequently, researchers will obtain (borrow) cell lines from other laboratories. While
this practice is widespread, it has one major drawback. There is a high probability that the
cells obtained in this manner will not be healthy, useful cultures. This is usually due to pre-
vious mix-ups or contamination with other cell lines, or the result of contamination with
microorganisms such as mycoplasmas, bacteria, fungi or yeast. These problems are covered
in detail in a Corning Technical Bulletin: Understanding and Managing Cell Culture
Contamination (Ref. 7).
What Are Cultured Cells Like?
Once in culture, cells exhibit a wide range of behaviors, characteristics and shapes. Some
of the more common ones are described below. John Paul discusses these issues in detail in
Chapter 3 of Cell and Tissue Culture (Ref. 3).
Corning culture dishes are
available in a variety of sizes
and shapes for growing
Cell Culture Systems
anchorage-dependent cells. Two basic culture systems are used for growing cells. These are based primarily upon the
ability of the cells to either grow attached to a glass or treated plastic substrate (Monolayer
Culture Sytems) or floating free in the culture medium (Suspension Culture Systems).
Monolayer cultures are usually grown in tissue culture treated dishes, T-flasks, roller bottles,
CellSTACK® Culture Chambers, or multiple well plates, the choice being based on the num-
ber of cells needed, the nature of the culture environment, cost and personal preference.
Suspension cultures are usually grown either:
1. In magnetically rotated spinner flasks or shaken Erlenmeyer flasks where the cells are
kept actively suspended in the medium;
2. In stationary culture vessels such as T-flasks and bottles where, although the cells are not
Corning culture flasks are kept agitated, they are unable to attach firmly to the substrate.
used for growing anchorage-
dependent cells. Many cell lines, especially those derived from normal tissues, are considered to be
Anchorage-Dependent, that is, they can only grow when attached to a suitable substrate.
Some cell lines that are no longer considered normal (frequently designated as Transformed
Cells) are frequently able to grow either attached to a substrate or floating free in suspension;
they are Anchorage-Independent. In addition, some normal cells, such as those found in
the blood, do not normally attach to substrates and always grow in suspension.
Types of Cells
Cultured cells are usually described based on their morphology (shape and appearance) or
their functional characteristics. There are three basic morphologies:
1. Epithelial-like: cells that are attached to a substrate and appear flattened and polygonal
in shape.
Corning spinner vessels are
used for growing anchorage- 2. Lymphoblast-like: cells that do not attach normally to a substrate but remain in
independent cells in suspension with a spherical shape.
suspension.
3. Fibroblast-like: cells that are attached to a substrate and appear elongated and bipolar,
frequently forming swirls in heavy cultures.
It is important to remember that the culture conditions play an important role in determin-
ing shape and that many cell cultures are capable of exhibiting multiple morphologies.
Using cell fusion techniques, it is also possible to obtain hybrid cells by fusing cells from
two different parents. These may exhibit characteristics of either parent or both parents.
This technique was used in 1975 to create cells capable of producing custom tailored mono-
clonal antibodies. These hybrid cells (called Hybridomas) are formed by fusing two differ-
ent but related cells. The first is a spleen-derived lymphocyte that is capable of producing
the desired antibody. The second is a rapidly dividing myeloma cell (a type of cancer cell)
Fibroblast-like 3T3 cells derived that has the machinery for making antibodies but is not programmed to produce any anti-
from mouse embryos
body. The resulting hybridomas can produce large quantities of the desired antibody. These
antibodies, called Monoclonal Antibodies due to their purity, have many important clini-
cal, diagnostic, and industrial applications with a yearly value of well over a billion dollars.
3
5. Anchorage-dependent cells also require a good substrate for attachment and growth. Glass
Basic environmental and specially treated plastics (to make the normally hydrophobic plastic surface hydrophilic
Requirements for or wettable) are the most commonly used substrates. However, Attachment Factors, such
“Happy” Cells: as collagen, gelatin, fibronectin and laminin, can be used as substrate coatings to improve
Q Controlled growth and function of normal cells derived from brain, blood vessels, kidney, liver, skin,
temperature etc. Often normal anchorage-dependent cells will also function better if they are grown on a
Q Good substrate for permeable or porous surface. This allows them to polarize (have a top and bottom through
cell attachment which things can enter and leave the cell) as they do in the body. Transwell® inserts are
Q Appropriate medium Corning vessels with membrane-based permeable supports that allow these cells to develop
and incubator that polarity and acquire the ability to exhibit special functions such as transport. Many special-
maintains the correct ized cells can only be truly “happy” (function normally) when grown on a porous substrate
pH and osmolality
in serum-free medium with the appropriate mixture of growth and attachment factors.
Cells can also be grown in suspension on beads made from glass, plastic, polyacrylamide and
cross-linked dextran molecules. This technique has been used to enable anchorage-dependent
cells to be grown in suspension culture systems and is increasingly important for the manu-
facture of cell-based biologicals.
The culture medium is the most important and complex factor to control in making cells
“happy”. Besides meeting the basic nutritional requirement of the cells, the culture medium
should also have any necessary growth factors, regulate the pH and osmolality, and provide
essential gases (O2 and CO2). The ‘food’ portion of the culture medium consists of amino
acids, vitamins, minerals, and carbohydrates. These allow the cells to build new proteins and
other components essential for growth and function as well as providing the energy neces-
Corning® Transwell permeable sary for metabolism. For additional information on this topic, see the article: Construction
supports are used to study cell
transport and migration.
of Tissue Culture Media by C. Waymouth in Growth, Nutrition and Metabolism of Cells in
Culture, Volume 1, (1972; Ref. 5).
The growth factors and hormones help regulate and control the cells’ growth rate and func-
tional characteristics. Instead of being added directly to the medium, they are often added in
an undefined manner by adding 5 to 20% of various animal sera to the medium. Unfortu-
nately, the types and concentration of these factors in serum vary considerably from batch
to batch. This often results in problems controlling growth and function. When growing
normal functional cells, sera are often replaced by specific growth factors.
The medium also controls the pH range of the culture and buffers the cells from abrupt
changes in pH. Usually a CO2-bicarbonate based buffer or an organic buffer, such as
HEPES, is used to help keep the medium pH in a range from 7.0 to 7.4 depending on the
type of cell being cultured. When using a CO2-bicarbonate buffer, it is necessary to regulate
the amount of CO2 dissolved in the medium. This is usually done using an incubator with
Suspension and microcarrier CO2 controls set to provide an atmosphere with between 2% and 10% CO2 (for Earle’s
cultures can be grown in glass salts-based buffers). However, some media use a CO2-bicarbonate buffer (for Hanks’ salts-
and plastic spinner vessels.
based buffers) that requires no additional CO2, but it must be used in a sealed vessel (not
dishes or plates). For additional information on this topic, see the article: The Gaseous
Environment of the Cell in Culture by W.F. McLimans in Growth, Nutrition and Metabolism
of Cells in Culture (1972; Ref. 5).
Finally, the osmolality (osmotic pressure) of the culture medium is important since it helps
regulate the flow of substances in and out of the cell. It is controlled by the addition or sub-
traction of salt in the culture medium. Evaporation of culture media from open culture ves-
sels (dishes, etc.) will rapidly increase the osmolality resulting in stressed, damaged or dead
cells. For open (not sealed) culture systems, incubators with high humidity levels to reduce
CHO-K1 cells growing on a evaporation are essential. For additional information, see article by C. Waymouth: Osmolality
microcarrier bead of Mammalian Blood and of Media for Culture of Mammalian Cells (1970; Ref. 6).
5
6. How to Decide if Cultured Cells Are “Happy”
Evaluating the general health or “happiness” of a culture is usually based on four important
cell characteristics: morphology, growth rate, plating efficiency and expression of special
functions. These same characteristics are also widely used in evaluating experimental results.
The Morphology or cell shape is the easiest to determine but is often the least useful. While
changes in morphology are frequently observed in cultures, it is often difficult to relate
these observations to the condition that caused them. It is also a very difficult characteristic
to quantify or to measure precisely.
Often, the first sign that something is wrong with a culture occurs when the cells are micro-
scopically examined and poor or unusual patterns of cell attachment or growth are observed.
When problems are suspected, staining the culture vessels with crystal violet or other simple
histological stains may show growth patterns indicating a problem. These growth problems
are discussed in detail in the Corning Technical Bulletin: General Guide for Identifying and
Correcting Common Cell Culture Growth and Attachment Problems (Ref. 8).
Examining the morphology of
these stained roller bottles Cell counting and other methods for estimating cell number, on the other hand, allow the
(containing MRC-5 human determination of the Growth Rate, which is sensitive to major changes in the culture
fibroblasts) is a good way to environment. This allows the design of experiments to determine which set of conditions
check if the cells are “happy”.
(culture media, substrate, serum, plasticware) is better
The bottle on the left was
rotated at too high a speed for the cells, i.e., the conditions producing the best growth rate. These same or similar
resulting in poor attachment techniques can also be used to measure cell survival or death and are often used for in vitro
and growth and very cytotoxicity assays.
“unhappy” cells.
Plating Efficiency is a testing method where small numbers of cells (20 to 200) are placed
in a culture vessel and the number of colonies they form is measured. The percentage of
cells forming colonies is a measure of survival, while the colony size is a measure of growth
rate. This testing method is similar in application to growth rate analysis but is more sensi-
tive to small variations in culture conditions.
The final characteristic, the Expression of Specialized Functions, is usually the most dif-
ficult to observe and measure. Usually biochemical or immunological assays and tests are
used. While cultured cells may grow very well in subobtimal conditions, highly specialized
functions usually require near perfect culture conditions and are often quickly lost when
cells are placed in culture.
A colony of fixed and stained
human fibroblast cells What is Cell Culture Used For?
Cell culture has become one of the major tools used in cell and molecular biology. Some of
the important areas where cell culture is currently playing a major role are briefly described
below:
Model Systems
Cell cultures provide a good model system for studying 1) basic cell biology and biochem-
istry, 2) the interactions between disease-causing agents and cells, 3) the effects of drugs on
cells, 4) the process and triggers for aging, and 5) nutritional studies.
Toxicity Testing
Cultured cells are widely used alone or in conjunction with animal tests to study the effects
of new drugs, cosmetics and chemicals on survival and growth in a wide variety of cell types.
Especially important are liver- and kidney-derived cell cultures.
Cancer Research
HTS Transwell®-24 plates are Since both normal cells and cancer cells can be grown in culture, the basic differences
used for toxicity testing and between them can be closely studied. In addition, it is possible, by the use of chemicals,
drug transport studies viruses and radiation, to convert normal cultured cells to cancer causing cells. Thus, the
mechanisms that cause the change can be studied. Cultured cancer cells also serve as a test
system to determine suitable drugs and methods for selectively destroying types of cancer.
6
7. Virology
One of the earliest and major uses of cell culture is the replication of viruses in cell cultures
(in place of animals) for use in vaccine production. Cell cultures are also widely used in the
clinical detection and isolation of viruses, as well as basic research into how they grow and
infect organisms.
Cell-Based Manufacturing
While cultured cells can be used to produce many important products, three areas are gen-
erating the most interest. The first is the large-scale production of viruses for use in vaccine
production. These include vaccines for polio, rabies, chicken pox, hepatitis B and measles.
Corning® roller bottles are Second, is the large-scale production of cells that have been genetically engineered to pro-
widely used for producing
viral vaccines.
duce proteins that have medicinal or commercial value. These include monoclonal antibodies,
insulin, hormones, etc. Third, is the use of cells as replacement tissues and organs. Artificial
skin for use in treating burns and ulcers is the first commercially available product. How-
ever, testing is underway on artificial organs such as pancreas, liver and kidney. A potential
supply of replacement cells and tissues may come out of work currently being done with both
embryonic and adult stem cells. These are cells that have the potential to differentiate into
a variety of different cell types. It is hoped that learning how to control the development of
these cells may offer new treatment approaches for a wide variety of medical conditions.
The Corning CellCube® Genetic Counseling
bioreactor system is ideal Amniocentesis, a diagnostic technique that enables doctors to remove and culture fetal cells
for mass production of from pregnant women, has given doctors an important tool for the early diagnosis of fetal
anchorage-dependent cells.
disorders. These cells can then be examined for abnormalities in their chromosomes and
genes using karyotyping, chromosome painting and other molecular techniques.
Genetic Engineering
The ability to transfect or reprogram cultured cells with new genetic material (DNA and
genes) has provided a major tool to molecular biologists wishing to study the cellular effects
of the expression of theses genes (new proteins). These techniques can also be used to pro-
duce these new proteins in large quantity in cultured cells for further study. Insect cells are
widely used as miniature cells factories to express substantial quantities of proteins that they
manufacture after being infected with genetically engineered baculoviruses.
Corning® Erlenmeyer flasks are
Gene Therapy
often used for growing insect The ability to genetically engineer cells has also led to their use for gene therapy. Cells can
cells in suspension. be removed from a patient lacking a functional gene and the missing or damaged gene can
then be replaced. The cells can be grown for a while in culture and then replaced into the
patient. An alternative approach is to place the missing gene into a viral vector and then
“infect’’ the patient with the virus in the hope that the missing gene will then be expressed
in the patient’s cells.
7
Drug Screening and Development
Cell-based assays have become increasingly important for the pharmaceutical industry, not
just for cytotoxicity testing but also for high throughput screening of compounds that may
have potential use as drugs. Originally, these cell culture tests were done in 96 well plates,
Corning microplates are widely but increasing use is now being made of 384 and 1536 well plates.
used for drug screening.
7