Subculturing
•
0 likes•635 views
Subculture involves transferring cells from an existing culture to fresh growth medium in order to prolong the life and expand the number of cells. This is necessary because over time toxic metabolites build up and nutrients are depleted in the original culture. Subculture produces a new culture with lower cell density, fresh nutrients and no toxins. It allows for both the long-term maintenance of cell lines and increasing cell numbers for industrial or research purposes. Different techniques are used depending on whether the cells are adherent or non-adherent.
Report
Share
Report
Share
Recommended
molecular marker AFLP, and application
Introduction
Steps in AFLP
Advantage of AFLP
Disadvantage of AFLP
Application
Conclusion
Reference
Types of cell culture
There are three main types of cell culture: primary cell culture, where cells are directly separated from tissue and grown; secondary cell culture, where primary cells are subcultured and transferred to new vessels; and cell lines, which can be either finite, with a limited lifespan, or continuous, capable of indefinite growth. Primary cell culture can involve adherent cells that attach to surfaces or suspension cells that float freely. Secondary culture involves detaching and transferring adherent cells to new vessels using enzymatic methods.
Aflp (amplified fragment length polymorphism), alu
Alu PCR amplifies repetitive Alu elements in DNA using Alu-specific primers. The presence or absence of Alu insertions can be used for population genetics, forensics, and paternity testing. Alu elements are short, inter
Bioreactors for animal cell suspension culture
This document discusses bioreactors for animal cell suspension culture. It begins by introducing animal cell culture and some key developments that enabled it. There are two main types of culture: primary culture using explants or enzymes, and secondary culture which is derived from primary culture. Cells can be anchorage-dependent, growing in monolayers, or non-anchorage dependent, growing in suspension. Bioreactors provide conditions for mass cultivation of suspension cells. Properties of animal cells require gentle mixing and aeration in bioreactors. Common bioreactor types for suspension culture include stirred tank, continuous flow, and airlift fermentors. Perfusion culture allows continuous medium exchange to achieve high cell densities and productivity.
Shuttle vector
1. Yeast plasmids like the 2 micron circle have been extensively studied and developed into yeast cloning vectors.
2. Shuttle vectors like YEp vectors contain selectable marker genes like LEU2 and bacterial plasmid origins of replication like pBR322, allowing them to replicate in both E. coli and yeast.
3. The 2 micron circle is a 6kb endogenous yeast plasmid that replicates autonomously through an ARS sequence and is maintained at 50-100 copies per cell.
Cytotoxicity
This document discusses cytotoxicity and methods for measuring it in vitro. It defines cytotoxicity as the ability of chemicals or cells to destroy living cells, which can lead to necrosis, apoptosis, or cytostasis. Measuring cytotoxicity is important for drug development and safety testing. In vitro assays are now commonly used as they are faster, cheaper, and more accurate than animal models. Several types of cytotoxicity assays are described, including dye exclusion assays like trypan blue, colorimetric assays like MTT, fluorometric assays like CFDA-AM, and luminometric assays like ATP assays. These assays measure endpoints such as membrane integrity, enzyme activity, proliferation, and ATP production to determine cytotoxic effects of chemicals on cells.
Primary culture and cell line
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
Cybrids
Cybrids are produced through the fusion of protoplasts from two different plant species, combining the cytoplasm of both but the nucleus of only one species. This technique allows for the transfer of cytoplasmic traits like male sterility between incompatible species. Protoplast isolation, fusion, selection, and regeneration of hybrid cells into whole plants are required to produce cybrids. Cybrids can be used to study cytoplasmic genes and transfer desirable agricultural traits, overcoming sexual incompatibility barriers in plant breeding.
Recommended
molecular marker AFLP, and application
Introduction
Steps in AFLP
Advantage of AFLP
Disadvantage of AFLP
Application
Conclusion
Reference
Types of cell culture
There are three main types of cell culture: primary cell culture, where cells are directly separated from tissue and grown; secondary cell culture, where primary cells are subcultured and transferred to new vessels; and cell lines, which can be either finite, with a limited lifespan, or continuous, capable of indefinite growth. Primary cell culture can involve adherent cells that attach to surfaces or suspension cells that float freely. Secondary culture involves detaching and transferring adherent cells to new vessels using enzymatic methods.
Aflp (amplified fragment length polymorphism), alu
Alu PCR amplifies repetitive Alu elements in DNA using Alu-specific primers. The presence or absence of Alu insertions can be used for population genetics, forensics, and paternity testing. Alu elements are short, inter
Bioreactors for animal cell suspension culture
This document discusses bioreactors for animal cell suspension culture. It begins by introducing animal cell culture and some key developments that enabled it. There are two main types of culture: primary culture using explants or enzymes, and secondary culture which is derived from primary culture. Cells can be anchorage-dependent, growing in monolayers, or non-anchorage dependent, growing in suspension. Bioreactors provide conditions for mass cultivation of suspension cells. Properties of animal cells require gentle mixing and aeration in bioreactors. Common bioreactor types for suspension culture include stirred tank, continuous flow, and airlift fermentors. Perfusion culture allows continuous medium exchange to achieve high cell densities and productivity.
Shuttle vector
1. Yeast plasmids like the 2 micron circle have been extensively studied and developed into yeast cloning vectors.
2. Shuttle vectors like YEp vectors contain selectable marker genes like LEU2 and bacterial plasmid origins of replication like pBR322, allowing them to replicate in both E. coli and yeast.
3. The 2 micron circle is a 6kb endogenous yeast plasmid that replicates autonomously through an ARS sequence and is maintained at 50-100 copies per cell.
Cytotoxicity
This document discusses cytotoxicity and methods for measuring it in vitro. It defines cytotoxicity as the ability of chemicals or cells to destroy living cells, which can lead to necrosis, apoptosis, or cytostasis. Measuring cytotoxicity is important for drug development and safety testing. In vitro assays are now commonly used as they are faster, cheaper, and more accurate than animal models. Several types of cytotoxicity assays are described, including dye exclusion assays like trypan blue, colorimetric assays like MTT, fluorometric assays like CFDA-AM, and luminometric assays like ATP assays. These assays measure endpoints such as membrane integrity, enzyme activity, proliferation, and ATP production to determine cytotoxic effects of chemicals on cells.
Primary culture and cell line
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
Cybrids
Cybrids are produced through the fusion of protoplasts from two different plant species, combining the cytoplasm of both but the nucleus of only one species. This technique allows for the transfer of cytoplasmic traits like male sterility between incompatible species. Protoplast isolation, fusion, selection, and regeneration of hybrid cells into whole plants are required to produce cybrids. Cybrids can be used to study cytoplasmic genes and transfer desirable agricultural traits, overcoming sexual incompatibility barriers in plant breeding.
Rapd ppt
The document describes a seminar on Random Amplified Polymorphic DNA (RAPD) markers. It defines RAPD as a type of PCR reaction that amplifies random segments of DNA using short arbitrary nucleotide primers. The document outlines the history, principle, procedure, applications, advantages, and limitations of RAPD analysis. It compares RAPD to other molecular marker techniques and concludes that RAPD is a lab technique used to amplify unknown DNA segments for analysis.
Introduction to animal cell culture
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.
Immobilized cell technology
This document discusses immobilized cell technology and its applications in the beer, wine, and dairy industries. It begins with an introduction to immobilization, which involves imprisoning cells or enzymes in a support or matrix. This allows cells to be reused as they are separated from products. The document then discusses specific applications of immobilized cell technology in wine production, beer production, and dairy industry production. It outlines various support materials and immobilization techniques used for each industry.
Constituent of animal tissue culture media and their specific application
INTRODUCTION
HISTORY
PHYSICOCHEMICAL PROPERTIES OF CULTURE MEDIA
pH
CO2, BICARBONATE AND BUFFERING
OXYGEN
TEMPERATURE
OSMOLALITY
BALANCED SALT SOLUTIONS
CONSTITUENTS OF CULTURE MEDIA
AMINO ACIDS
VITAMINS
SALTS
GLUCOSE
OTHER ORGANIC SUPPLEMENTS
ANTIBIOTICS
SERUM
PROTEINS
NUTRIENTS AND METABOLITES
HORMONES AND GROWTH FACTORS
LIPIDS
MINERALS
INHIBITORS
APPLICATIONS OF CULTURE MEDIA
CONCLUSION
REFERENCES
Somatic embryogenesis, in plant tissue culture
INTRODUCTION
HISTORY
SOMATIC EMBRYOGENESIS DEVELOPMENT
- INDUCTION
- DEVELOPMENT
- MATURATION
SYNTHETIC SEED
FACTORS AFFECTING SOMATIC EMBRYOGENESIS
APPLICATIONS
REFERENCE
AFLP, RFLP & RAPD
Techniques based on the principle of selectively amplifying a subset of restriction fragments from a complex mixture of DNA fragments obtained after digestion of genomic DNA with restriction endonucleases.
Morphological, Cytological and Biochemical Markers
I've put a lot of effort for create this presentation. This'll help to lot of biotechnology and agricultural students for there assignments and exam study.
Agrose gel electrophoresis
This document summarizes the process of agarose gel electrophoresis. Agarose gel is prepared by combining agarose powder with a buffer solution to establish pH and conductivity. Samples are loaded into wells in the gel and an electric current is applied, causing DNA fragments to migrate through the gel at rates corresponding to their size. Agarose gel electrophoresis is used to separate and analyze DNA fragments for applications such as estimating DNA size, analyzing polymerase chain reaction products, and extracting DNA fragments for further purification.
Development of inoculum buildup
This document discusses the development of inoculum for industrial fermentation processes. It defines inoculum as a mixture of cultured microbes and the media they are growing in. The key steps in inoculum development are preparing a suitable growth media, maintaining optimal pH and nutrient levels, and conducting growth in stepwise increasing volumes. Examples of common inoculum media compositions are provided for vitamin and bacterial insecticide production processes. Developing high quality inoculum is important for efficiently adapting cultures to fermentation conditions.
Lipases & Industrial Production of Lipases
Introduction to Lipases
Industrial Production of Lipases by Fermentation
Industrial Significance of Lipases
Cell suspension culture
This document discusses cell suspension culture, which involves growing plant cells, tissues, or organs in liquid nutrient medium. There are two main types of cell suspension cultures: batch culture and continuous culture. Batch culture involves growing cells in a finite volume of agitated medium, while continuous culture replaces old medium continuously to maintain cells in steady growth. The document outlines different methods for each type of culture and their characteristics, advantages, and importance for studying plant cell physiology and biochemistry.
Shuttle vector - a plasmid vector used in rDNA technology.
Shuttle vectors are constructed so that they can propagate in both eukaryotic and prokaryotic cells. They contain two origins of replication - one for each host cell type. YEp13 is an example of a shuttle vector that can replicate in yeast (eukaryotic) and E. coli (prokaryotic) cells. It contains sequences from the 2 micron yeast plasmid, including the LEU2 gene as a selectable marker, as well as sequences from the pBR322 plasmid which allow replication in E. coli. Shuttle vectors allow initial cloning experiments and selection to be done in E. coli before introducing the recombinant DNA into yeast cells.
Establishment and maintenance of callus and suspension culture.pptx
This document discusses callus and suspension cultures. Callus cultures involve growing loose aggregates of parenchyma cells on a solid nutrient medium. Suspension cultures grow tissues and cells in liquid medium with constant agitation. Batch cultures use a limited supply of nutrients until they are depleted, while continuous cultures maintain a steady state by draining out used medium and adding fresh medium. Both callus and suspension cultures need to be sub-cultured regularly to maintain healthy growth.
Western blotting
The western blot is a technique used to detect specific proteins in a sample. It involves separating proteins by size using gel electrophoresis, transferring them to a membrane, and using antibodies to detect the target protein. The key steps are sample preparation, gel electrophoresis, blotting, blocking, antibody probing, and detection. Western blotting allows researchers to identify proteins from complex mixtures and is widely used in molecular biology and medical diagnosis, such as detecting HIV, HBV, and HSV infections.
Equipment and materials for animal cell culture
INTRODUCTION
REQUIREMENTS OF A TISSUE CULTURE LABORATORY
TISSUE CULTURE EQUIPMENTS AND MATERIALS
PREPARATION AND STERILIZATION
SUPPLEMENTARY LABORATORY EQUIPMENT
SPECIALIZED EQUIPMENT
CONCLUSION
REFRENCE
Types of animal cell culture; characterization & Their preservation.
This document provides an overview of animal cell culture, including the different types (primary and secondary cell culture, cell lines), techniques used for primary culture, and characterization and preservation of animal cells. It discusses how primary cell culture involves separating cells directly from tissue and allowing them to grow under controlled conditions. Secondary cell culture refers to sub-culturing primary cells by transferring them to new vessels with fresh media. Cell lines can be propagated repeatedly and sometimes indefinitely. The document also describes cryopreservation as a method for preserving live cells and tissues at ultra-low temperatures in liquid nitrogen.
Subculturing
Subculture involves transferring cells from an existing culture to fresh growth medium in order to prolong the life and expand the number of cells. This is necessary because over time toxic metabolites build up and nutrients are depleted in the original culture. Subculture produces a new culture with lower cell density and fresh nutrients. It allows for long-term maintenance of cell lines and increasing cell numbers for industrial or research purposes. Different techniques are used to subculture adherent versus non-adherent cells.
Characterization of cell line by Cell morphology, Chromosome analysis and DNA...
The document discusses various methods for characterizing cell lines, including morphology, chromosome analysis, and DNA content. Morphology examines cell size, shape, and structure to classify cell lines as fibroblastic, epithelial-like, lymphoblast-like, endothelial, or neuronal. Chromosome analysis techniques like karyotyping, banding, and fluorescence in situ hybridization are used to identify species, detect genetic abnormalities, and confirm identity. DNA content analysis using stains like propidium iodide measure ploidy and identify transformed cells. Characterization is essential for confirming cell line identity and detecting contamination or genetic instability.
Restriction Mapping
Restriction enzymes cut DNA molecules at specific recognition sites. Restriction mapping involves digesting an unknown DNA segment with restriction enzymes and analyzing the fragment sizes to determine the locations of restriction sites. One method involves single and double digestions with two enzymes followed by gel electrophoresis to separate the fragments by size. By comparing the fragment patterns between single and double digestions, the positions of each restriction site can be mapped, generating a restriction map of the DNA segment. Restriction mapping was previously important for characterizing cloned DNA but is now easier using DNA sequencing, though analysis of restriction sites remains useful for comparing chromosomal organization between strains.
Explant sterilization
This document provides instructions for sterilizing plant explants for tissue culture. It discusses that culture media supports microbial growth which can kill plant tissues, so complete sterilization is essential. The key steps outlined are: washing explants with detergent and water; submerging in disinfectants like ethanol or mercury chloride; rinsing with sterile water; and sealing cultures with sterile cotton plugs. A two-step sterilization using ethanol prior to another disinfectant can improve sterility. Proper sterilization will increase the success rate of tissue culture by reducing contamination and allowing explant growth without microbes.
Ubaid afzal (10)
Subculturing involves transferring cells from a previous culture into fresh growth medium in order to propagate the cell strain, prolong the life of the culture, and expand the number of cells. It is important for both proliferating and non-proliferating cell types. The protocol for subculturing depends on whether the cells are adherent or non-adherent. For adherent cells, trypsin is used to detach the cells before transferring them to fresh medium and seeding new culture vessels.
Animal cell culture (Basics)
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.
More Related Content
What's hot
Rapd ppt
The document describes a seminar on Random Amplified Polymorphic DNA (RAPD) markers. It defines RAPD as a type of PCR reaction that amplifies random segments of DNA using short arbitrary nucleotide primers. The document outlines the history, principle, procedure, applications, advantages, and limitations of RAPD analysis. It compares RAPD to other molecular marker techniques and concludes that RAPD is a lab technique used to amplify unknown DNA segments for analysis.
Introduction to animal cell culture
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.
Immobilized cell technology
This document discusses immobilized cell technology and its applications in the beer, wine, and dairy industries. It begins with an introduction to immobilization, which involves imprisoning cells or enzymes in a support or matrix. This allows cells to be reused as they are separated from products. The document then discusses specific applications of immobilized cell technology in wine production, beer production, and dairy industry production. It outlines various support materials and immobilization techniques used for each industry.
Constituent of animal tissue culture media and their specific application
INTRODUCTION
HISTORY
PHYSICOCHEMICAL PROPERTIES OF CULTURE MEDIA
pH
CO2, BICARBONATE AND BUFFERING
OXYGEN
TEMPERATURE
OSMOLALITY
BALANCED SALT SOLUTIONS
CONSTITUENTS OF CULTURE MEDIA
AMINO ACIDS
VITAMINS
SALTS
GLUCOSE
OTHER ORGANIC SUPPLEMENTS
ANTIBIOTICS
SERUM
PROTEINS
NUTRIENTS AND METABOLITES
HORMONES AND GROWTH FACTORS
LIPIDS
MINERALS
INHIBITORS
APPLICATIONS OF CULTURE MEDIA
CONCLUSION
REFERENCES
Somatic embryogenesis, in plant tissue culture
INTRODUCTION
HISTORY
SOMATIC EMBRYOGENESIS DEVELOPMENT
- INDUCTION
- DEVELOPMENT
- MATURATION
SYNTHETIC SEED
FACTORS AFFECTING SOMATIC EMBRYOGENESIS
APPLICATIONS
REFERENCE
AFLP, RFLP & RAPD
Techniques based on the principle of selectively amplifying a subset of restriction fragments from a complex mixture of DNA fragments obtained after digestion of genomic DNA with restriction endonucleases.
Morphological, Cytological and Biochemical Markers
I've put a lot of effort for create this presentation. This'll help to lot of biotechnology and agricultural students for there assignments and exam study.
Agrose gel electrophoresis
This document summarizes the process of agarose gel electrophoresis. Agarose gel is prepared by combining agarose powder with a buffer solution to establish pH and conductivity. Samples are loaded into wells in the gel and an electric current is applied, causing DNA fragments to migrate through the gel at rates corresponding to their size. Agarose gel electrophoresis is used to separate and analyze DNA fragments for applications such as estimating DNA size, analyzing polymerase chain reaction products, and extracting DNA fragments for further purification.
Development of inoculum buildup
This document discusses the development of inoculum for industrial fermentation processes. It defines inoculum as a mixture of cultured microbes and the media they are growing in. The key steps in inoculum development are preparing a suitable growth media, maintaining optimal pH and nutrient levels, and conducting growth in stepwise increasing volumes. Examples of common inoculum media compositions are provided for vitamin and bacterial insecticide production processes. Developing high quality inoculum is important for efficiently adapting cultures to fermentation conditions.
Lipases & Industrial Production of Lipases
Introduction to Lipases
Industrial Production of Lipases by Fermentation
Industrial Significance of Lipases
Cell suspension culture
This document discusses cell suspension culture, which involves growing plant cells, tissues, or organs in liquid nutrient medium. There are two main types of cell suspension cultures: batch culture and continuous culture. Batch culture involves growing cells in a finite volume of agitated medium, while continuous culture replaces old medium continuously to maintain cells in steady growth. The document outlines different methods for each type of culture and their characteristics, advantages, and importance for studying plant cell physiology and biochemistry.
Shuttle vector - a plasmid vector used in rDNA technology.
Shuttle vectors are constructed so that they can propagate in both eukaryotic and prokaryotic cells. They contain two origins of replication - one for each host cell type. YEp13 is an example of a shuttle vector that can replicate in yeast (eukaryotic) and E. coli (prokaryotic) cells. It contains sequences from the 2 micron yeast plasmid, including the LEU2 gene as a selectable marker, as well as sequences from the pBR322 plasmid which allow replication in E. coli. Shuttle vectors allow initial cloning experiments and selection to be done in E. coli before introducing the recombinant DNA into yeast cells.
Establishment and maintenance of callus and suspension culture.pptx
This document discusses callus and suspension cultures. Callus cultures involve growing loose aggregates of parenchyma cells on a solid nutrient medium. Suspension cultures grow tissues and cells in liquid medium with constant agitation. Batch cultures use a limited supply of nutrients until they are depleted, while continuous cultures maintain a steady state by draining out used medium and adding fresh medium. Both callus and suspension cultures need to be sub-cultured regularly to maintain healthy growth.
Western blotting
The western blot is a technique used to detect specific proteins in a sample. It involves separating proteins by size using gel electrophoresis, transferring them to a membrane, and using antibodies to detect the target protein. The key steps are sample preparation, gel electrophoresis, blotting, blocking, antibody probing, and detection. Western blotting allows researchers to identify proteins from complex mixtures and is widely used in molecular biology and medical diagnosis, such as detecting HIV, HBV, and HSV infections.
Equipment and materials for animal cell culture
INTRODUCTION
REQUIREMENTS OF A TISSUE CULTURE LABORATORY
TISSUE CULTURE EQUIPMENTS AND MATERIALS
PREPARATION AND STERILIZATION
SUPPLEMENTARY LABORATORY EQUIPMENT
SPECIALIZED EQUIPMENT
CONCLUSION
REFRENCE
Types of animal cell culture; characterization & Their preservation.
This document provides an overview of animal cell culture, including the different types (primary and secondary cell culture, cell lines), techniques used for primary culture, and characterization and preservation of animal cells. It discusses how primary cell culture involves separating cells directly from tissue and allowing them to grow under controlled conditions. Secondary cell culture refers to sub-culturing primary cells by transferring them to new vessels with fresh media. Cell lines can be propagated repeatedly and sometimes indefinitely. The document also describes cryopreservation as a method for preserving live cells and tissues at ultra-low temperatures in liquid nitrogen.
Subculturing
Subculture involves transferring cells from an existing culture to fresh growth medium in order to prolong the life and expand the number of cells. This is necessary because over time toxic metabolites build up and nutrients are depleted in the original culture. Subculture produces a new culture with lower cell density and fresh nutrients. It allows for long-term maintenance of cell lines and increasing cell numbers for industrial or research purposes. Different techniques are used to subculture adherent versus non-adherent cells.
Characterization of cell line by Cell morphology, Chromosome analysis and DNA...
The document discusses various methods for characterizing cell lines, including morphology, chromosome analysis, and DNA content. Morphology examines cell size, shape, and structure to classify cell lines as fibroblastic, epithelial-like, lymphoblast-like, endothelial, or neuronal. Chromosome analysis techniques like karyotyping, banding, and fluorescence in situ hybridization are used to identify species, detect genetic abnormalities, and confirm identity. DNA content analysis using stains like propidium iodide measure ploidy and identify transformed cells. Characterization is essential for confirming cell line identity and detecting contamination or genetic instability.
Restriction Mapping
Restriction enzymes cut DNA molecules at specific recognition sites. Restriction mapping involves digesting an unknown DNA segment with restriction enzymes and analyzing the fragment sizes to determine the locations of restriction sites. One method involves single and double digestions with two enzymes followed by gel electrophoresis to separate the fragments by size. By comparing the fragment patterns between single and double digestions, the positions of each restriction site can be mapped, generating a restriction map of the DNA segment. Restriction mapping was previously important for characterizing cloned DNA but is now easier using DNA sequencing, though analysis of restriction sites remains useful for comparing chromosomal organization between strains.
Explant sterilization
This document provides instructions for sterilizing plant explants for tissue culture. It discusses that culture media supports microbial growth which can kill plant tissues, so complete sterilization is essential. The key steps outlined are: washing explants with detergent and water; submerging in disinfectants like ethanol or mercury chloride; rinsing with sterile water; and sealing cultures with sterile cotton plugs. A two-step sterilization using ethanol prior to another disinfectant can improve sterility. Proper sterilization will increase the success rate of tissue culture by reducing contamination and allowing explant growth without microbes.
What's hot (20)
Constituent of animal tissue culture media and their specific application
Constituent of animal tissue culture media and their specific application
Morphological, Cytological and Biochemical Markers
Morphological, Cytological and Biochemical Markers
Shuttle vector - a plasmid vector used in rDNA technology.
Shuttle vector - a plasmid vector used in rDNA technology.
Establishment and maintenance of callus and suspension culture.pptx
Establishment and maintenance of callus and suspension culture.pptx
Types of animal cell culture; characterization & Their preservation.
Types of animal cell culture; characterization & Their preservation.
Characterization of cell line by Cell morphology, Chromosome analysis and DNA...
Characterization of cell line by Cell morphology, Chromosome analysis and DNA...
Similar to Subculturing
Ubaid afzal (10)
Subculturing involves transferring cells from a previous culture into fresh growth medium in order to propagate the cell strain, prolong the life of the culture, and expand the number of cells. It is important for both proliferating and non-proliferating cell types. The protocol for subculturing depends on whether the cells are adherent or non-adherent. For adherent cells, trypsin is used to detach the cells before transferring them to fresh medium and seeding new culture vessels.
Animal cell culture (Basics)
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.
SHS.320.Lec-10...pptx Virology & parasitology
This document provides information about the course CODE 320 on Virology/Parasitology. It discusses cell culture, including its history, terminology, techniques, types of cells, equipment used, and factors that affect cell growth. Key points include that cell culture involves growing cells outside their natural environment, the importance of subculture when cells reach confluence, and that cell growth follows distinct lag, log/exponential, and plateau phases.
PHMD202-Growth-of-Bacteria_144413.pptx
Bacteria reproduce through binary fission, where a single cell divides into two daughter cells. This results in exponential growth of the bacterial population over time. A bacterial growth curve typically shows four phases: a lag phase as the bacteria adapt, an exponential or log phase of rapid growth, a stationary phase as resources are depleted, and a death phase. Continuous culturing techniques like chemostats can maintain bacteria in the log phase of growth through controlled dilution of the culture medium.
Growth curve of bacteria
1. Bacteria growing in batch culture produce a growth curve with up to four distinct phases: lag, log, stationary, and death.
2. During the lag phase, bacteria are adjusting to their new medium and preparing for active growth.
3. The log phase is when bacteria grow exponentially as cell division occurs rapidly under favorable conditions.
4. As nutrients become depleted and waste accumulates, bacteria enter the stationary phase where growth stops.
5. In the death phase, cell numbers decline as bacteria can no longer survive due to nutrient depletion and waste accumulation.
METHODS AND APPLICATIONS OF TISSUE CULTURE.pptx
Tissue culture is a method of biological research where plant tissues are grown under sterile conditions. There are several methods of plant tissue culture including seed culture, embryo culture, callus culture, cell culture, bud culture, meristem culture, and protoplast culture. Each method involves explants from various plant tissues being placed on nutrient media to induce cell growth and differentiation. Tissue culture has many applications including rapid clonal propagation, inducing genetic variation, producing transgenic plants, and conserving plant genetic resources.
Animal cell culture
This presentation covers introduction,cell culture media,types of culture media, type of cell culture, common procedure and applications
immobilized cell reactor
This document discusses different methods for immobilizing whole cells, including perfusion bioreactors and biofilm formation. Perfusion bioreactors culture cells continuously over long periods by feeding fresh media and removing waste, while various separation methods like hollow fiber membranes or centrifuges keep cells in the bioreactor. Perfusion offers advantages like improved product quality, smaller reactor size, and lower costs compared to traditional fed-batch systems. The document also covers immobilizing cells through entrapment in polymers, attachment to surfaces, or passive biofilm formation on supports.
Ubaid afzal (3)
Sub-culturing involves transferring cells from an existing culture to fresh growth medium in order to prolong the life of the culture, expand the number of cells, and provide fresh nutrients and remove toxic metabolites. It is important for both proliferating and non-proliferating cell lines to prevent cell death. There are four types of sub-culturing: solid to solid, solid to liquid, liquid to solid, and liquid to liquid, which involve transferring microorganisms between agar plates, broths, or between the two types of media. Proper labeling of containers is important for identification.
Batch ,continous and synchronus culture.pptx
Defination,growth curve, types and kinetics of growth curve, applications and advantages and disadvantages . Environmental factors affecting the cell growth.
Applications of fish celllines by B.pptx
Recent research studies on fish cell lines found many more application of cell lines pathological studies, toxicology, biomedical research, vaccine development etc
Basic of Cell Culture (1).ppt
Cell culture is the process of growing cells under controlled conditions outside of their natural environment. Some key developments in cell culture technology include the use of antibiotics to prevent contamination, trypsin to detach adherent cells for subculturing, and chemically defined culture media. Cell culture is used for a variety of purposes like studying cell biology, testing drug toxicity, cancer research, virology, genetic engineering, and gene therapy. Proper aseptic techniques and safety precautions must be followed when working with cell cultures.
Physiology of microorganism 1
Microorganisms can reproduce through binary fission or budding. Their growth follows four phases: lag, exponential, stationary, and death. Key factors that affect microbial growth are nutrients, temperature, pH, moisture, oxygen, and osmotic potential. Growth is measured directly by counting colonies, cells, or turbidity or indirectly by metabolic activity or mass determination. Microorganisms are controlled through sterilization, disinfection, sanitization, antiseptics, chemotherapy, filtration, and radiation. Sterilization kills all microbes while disinfection and sanitization reduce microbes to safe levels.
Bacterial Growth and structure in book of bacteriology
This document summarizes bacterial growth and methods for measuring it. It describes the stages of bacterial growth (lag phase, exponential phase, stationary phase, death phase) and the process of binary fission that bacteria use to replicate. It also discusses two common ways to measure bacterial growth: microscopic cell counts, which involve direct observation of cells under a microscope, and turbidity measurements, which measure cell mass indirectly. Microscopic counts have limitations like difficulty distinguishing live from dead cells and achieving precision.
Dr. Rehab Al Mousa. Plant Tissue Culture
Plant tissue culture is a technique for growing plant cells, tissues or organs in vitro on artificial nutrient media under sterile conditions. It allows for the clonal propagation of plants as well as applications in plant breeding such as haploid production, somatic hybridization and genetic modification. Some challenges include contamination, hyperhydricity, phenolic exudation, shoot tip necrosis and somaclonal variation. Tissue culture has many uses in micropropagation, plant breeding, germplasm preservation, plant physiology and production of secondary metabolites.
Histotypic culture
Histotypic culture involves growing cell lines in a three-dimensional matrix at high density to form tissue-like structures. Common techniques include using gels, sponges, hollow fibers, spheroids, and rotating chambers to provide a 3D environment that allows cells to organize similarly to how they would in tissues. This allows for the study of processes like drug penetration and cell differentiation that are not possible with traditional 2D cultures. While histotypic cultures provide a model for certain tissue functions, they also face challenges like loss of cell differentiation over time.
cell_cutlure_0.pptx
Cell culture involves growing cells under controlled conditions outside of their natural environment. There are several types of culture, including organ culture, tissue culture, and cell culture. Cell culture allows cells to be studied without the complexity of an entire organism and provides a model for understanding cell behavior. Key factors that must be controlled include the culture surface, growth media formulation, temperature, gas exchange, and passaging of cells to maintain healthy growth.
Growth by Dr. Pandian M.
Microbial growth involves an increase in cell number through cell division. There are four phases of bacterial growth in a batch culture: lag phase, exponential/log phase, stationary phase, and death phase. During exponential phase, the population doubles in each generation time. Environmental factors like temperature, pH, oxygen levels, and nutrients influence microbial growth rates.
Animal Cell Culture.pptx
Presentation on Animal Cell Culture. Different types, requirements, advantages and disadvantages etc.
Similar to Subculturing (20)
Bacterial Growth and structure in book of bacteriology
Bacterial Growth and structure in book of bacteriology
More from BOTANYWith
Practical notebook botany paper A
Botany students taking Paper A will need to be prepared to demonstrate their knowledge of botany earned through either a Bachelor of Science (BS) or Bachelor of Science (BSc) degree program. The paper examines core topics in the field of botany and botanical sciences. Students should review their textbooks, class notes, and materials from relevant botany courses to prepare for Paper A.
Coronavirus diease, Symptom & Prevention
The document discusses the benefits of exercise for mental health. Regular physical activity can help reduce anxiety and depression and improve mood and cognitive functioning. Exercise boosts blood flow, releases endorphins, and promotes changes in the brain which help enhance one's emotional well-being and mental clarity.
Mitosis & Meioses
The document discusses the benefits of exercise for mental health. Regular physical activity can help reduce anxiety and depression and improve mood and cognitive function. Exercise causes chemical changes in the brain that may help protect against mental illness and improve symptoms.
Leaf, stomata & stem anatomy
The document discusses the benefits of exercise for mental health. Regular physical activity can help reduce anxiety and depression and improve mood and cognitive function. Exercise causes chemical changes in the brain that may help protect against mental illness and improve symptoms.
Bryophyte presentation
Bryophyte refers to non-vascular land plants that reproduce via spores and include about 25,000 species worldwide. They are found in many habitats and surfaces around the world. Bryophytes do not have true roots but have root-like rhizoids, and they are gametophyte dominant with the sporophyte remaining attached to and dependent on the gametophyte.
PCR
PCR is a method for amplifying DNA that allows one molecule to be increased to thousands within hours using DNA polymerase and building blocks. It has many roles including forensic applications, infectious disease diagnosis, medical applications, and research. PCR can fail due to contamination, so techniques have been developed for optimization including dedicating lab areas, adding DMSO to minimize DNA folding, determining the optimal magnesium concentration as a cofactor for polymerase, and using disposable equipment and thorough cleaning to prevent contamination between reactions.
Stem
Monocot stems have vascular bundles near the outer edge surrounded by large parenchyma cells, with no pith region. Dicot stems have vascular bundles in a ring around a central pith region made of parenchyma cells. Secondary growth occurs via cell division in cambium layers, thickening stems and roots, while primary growth happens at tips through cell division, elongating stems and roots with primary tissue.
Roots
The document discusses the benefits of exercise for mental health. Regular physical activity can help reduce anxiety and depression and improve mood and cognitive function. Exercise causes chemical changes in the brain that may help protect against developing mental illness and improve symptoms for those who already suffer from conditions like anxiety and depression.
Angiosperm
All living things go through similar stages of growth and development. They are born, mature into adults, reproduce, and eventually die. The series of changes that an organism goes through from its beginning as an offspring to its mature form is known as its life cycle.
Pteropsida
The document discusses the benefits of exercise for mental health. Regular physical activity can help reduce anxiety and depression and improve mood and cognitive functioning. Exercise boosts blood flow, releases endorphins, and promotes changes in the brain which help enhance one's emotional well-being and mental clarity.
Algae
Algae are classified into taxonomic groups using the same rules as land plants, but the organization of algal groups above the order level has changed significantly since 1960.
Fungus
Fungi were once classified as plants but are actually very different, belonging to their own kingdom. Unlike plants, fungi do not have chlorophyll or photosynthesize. They obtain nutrients by decomposing and breaking down organic material like dead plants and animals through long branching tubes called hyphae that can cover large areas.
Virus
Viruses are small infectious agents that can only replicate inside living cells. They infect all forms of life from animals and plants to bacteria. Louis Pasteur first speculated about pathogens too small to be seen under microscopes to cause diseases like rabies. The first images of viruses were obtained after the invention of electron microscopes in 1931. Viruses come in different shapes and sizes and infect cells by hijacking their replication machinery to produce more viruses.
Basics of molecular biology tools and techniques
The key players in molecular biology are DNA, RNA, and proteins. DNA is the blueprint stored in the genome that contains the genetic instructions. It is replicated for cell division. During transcription, a complementary RNA copy of a DNA sequence is generated. There are several types of RNA including mRNA and rRNA. mRNA is translated by ribosomes into proteins, the functional molecules that carry out most tasks in cells. Various techniques are used in molecular biology like PCR, gel electrophoresis, and blotting to study these biomolecules.
Ppt
Water logging occurs when excess rainfall or high water tables prevent soils from adequately draining water, reducing oxygen levels in the soil. In Pakistan, 38% of irrigated land is damaged by water logging, with 14% also affected by salinity. Water logging primarily impacts agriculture, with an estimated 50% of land in Pakistan affected and 30 million acres rendered uncultivated. It can prevent plant germination and growth, cause nutrient deficiencies, and even kill plants. Solutions include planting trees and bushes to control water tables, installing drainage systems, and choosing appropriate crops, seedlings, fertilization, and weed control.
Study of Transcriptome
Transcriptomics is the study of the transcriptome, which is the complete set of RNA transcripts produced by the genome under certain conditions, using high-throughput methods like microarray analysis. The transcriptome includes mRNA, rRNA, tRNA and other non-coding RNA transcribed in a cell or population of cells. Oncogenomics applies transcriptomics to characterize genes associated with cancer. Gene expression analysis focuses on relevant target genes and their location and distances on chromosomes can be determined through sequence mapping. Non-coding RNAs regulate gene expression at the transcriptional and post-transcriptional levels.
Success stories.
This document discusses success stories of developing disease-resistant plant varieties through conventional breeding and genetic engineering techniques. It provides examples of transgenic crops with enhanced traits like increased yield, nutritional quality, stress tolerance and pest resistance. Specifically, it mentions golden rice which was engineered to produce beta-carotene, and virus-resistant plants containing viral coat proteins. The document also discusses delayed fruit ripening tomatoes and insect-resistant Bt crops. Overall the document outlines major advances in developing stress-tolerant and nutritionally-enhanced transgenic crops.
Use of legumes
This document discusses using legume-microbe symbioses for phytoremediation of heavy metal polluted soils. Legumes form symbiotic relationships with arbuscular mycorrhizal fungi, plant growth-promoting rhizobacteria, and endophytic microorganisms which help the plants grow in contaminated soils. Mono-inoculation or combined inoculation with different microorganisms can increase legume growth and uptake of heavy metals. However, accelerated heavy metal uptake may cause toxic effects or inhibit plant growth. The interactions between heavy metal tolerance of symbiotic plant and microbe partners needs to be better understood.
Ppt
Vacuoles play several important roles in plant cells. They can contain multiple compartments to carry out different functions. Transporters in the vacuolar membrane, including proton pumps, aquaporins, and various channels, regulate the transport of substances like sugars, acids, and secondary metabolites into and out of the vacuole. This transport affects processes like fruit growth, taste, and quality. Proton pumps generate a proton gradient across the membrane that drives secondary active transport. Aquaporins transport water, while channels transport sugars, organic acids, and other compounds to store or sequester them in the vacuole.
More from BOTANYWith (20)
Recently uploaded
Solutons Maths Escape Room Spatial .pptx
Solutions of Puzzles of Mathematics Escape Room Game in Spatial.io
How to Setup Warehouse & Location in Odoo 17 Inventory
In this slide, we'll explore how to set up warehouses and locations in Odoo 17 Inventory. This will help us manage our stock effectively, track inventory levels, and streamline warehouse operations.
ANATOMY AND BIOMECHANICS OF HIP JOINT.pdf
it describes the bony anatomy including the femoral head , acetabulum, labrum . also discusses the capsule , ligaments . muscle that act on the hip joint and the range of motion are outlined. factors affecting hip joint stability and weight transmission through the joint are summarized.
BÀI TẬP BỔ TRỢ TIẾNG ANH LỚP 9 CẢ NĂM - GLOBAL SUCCESS - NĂM HỌC 2024-2025 - ...
BÀI TẬP BỔ TRỢ TIẾNG ANH LỚP 9 CẢ NĂM - GLOBAL SUCCESS - NĂM HỌC 2024-2025 - ...Nguyen Thanh Tu Collection
https://app.box.com/s/tacvl9ekroe9hqupdnjruiypvm9rdaneAdvanced Java[Extra Concepts, Not Difficult].docx
This is part 2 of my Java Learning Journey. This contains Hashing, ArrayList, LinkedList, Date and Time Classes, Calendar Class and more.
How to deliver Powerpoint Presentations.pptx
"How to make and deliver dynamic presentations by making it more interactive to captivate your audience attention"
Chapter wise All Notes of First year Basic Civil Engineering.pptx
Chapter wise All Notes of First year Basic Civil Engineering
Syllabus
Chapter-1
Introduction to objective, scope and outcome the subject
Chapter 2
Introduction: Scope and Specialization of Civil Engineering, Role of civil Engineer in Society, Impact of infrastructural development on economy of country.
Chapter 3
Surveying: Object Principles & Types of Surveying; Site Plans, Plans & Maps; Scales & Unit of different Measurements.
Linear Measurements: Instruments used. Linear Measurement by Tape, Ranging out Survey Lines and overcoming Obstructions; Measurements on sloping ground; Tape corrections, conventional symbols. Angular Measurements: Instruments used; Introduction to Compass Surveying, Bearings and Longitude & Latitude of a Line, Introduction to total station.
Levelling: Instrument used Object of levelling, Methods of levelling in brief, and Contour maps.
Chapter 4
Buildings: Selection of site for Buildings, Layout of Building Plan, Types of buildings, Plinth area, carpet area, floor space index, Introduction to building byelaws, concept of sun light & ventilation. Components of Buildings & their functions, Basic concept of R.C.C., Introduction to types of foundation
Chapter 5
Transportation: Introduction to Transportation Engineering; Traffic and Road Safety: Types and Characteristics of Various Modes of Transportation; Various Road Traffic Signs, Causes of Accidents and Road Safety Measures.
Chapter 6
Environmental Engineering: Environmental Pollution, Environmental Acts and Regulations, Functional Concepts of Ecology, Basics of Species, Biodiversity, Ecosystem, Hydrological Cycle; Chemical Cycles: Carbon, Nitrogen & Phosphorus; Energy Flow in Ecosystems.
Water Pollution: Water Quality standards, Introduction to Treatment & Disposal of Waste Water. Reuse and Saving of Water, Rain Water Harvesting. Solid Waste Management: Classification of Solid Waste, Collection, Transportation and Disposal of Solid. Recycling of Solid Waste: Energy Recovery, Sanitary Landfill, On-Site Sanitation. Air & Noise Pollution: Primary and Secondary air pollutants, Harmful effects of Air Pollution, Control of Air Pollution. . Noise Pollution Harmful Effects of noise pollution, control of noise pollution, Global warming & Climate Change, Ozone depletion, Greenhouse effect
Text Books:
1. Palancharmy, Basic Civil Engineering, McGraw Hill publishers.
2. Satheesh Gopi, Basic Civil Engineering, Pearson Publishers.
3. Ketki Rangwala Dalal, Essentials of Civil Engineering, Charotar Publishing House.
4. BCP, Surveying volume 1
Hindi varnamala | hindi alphabet PPT.pdf
हिंदी वर्णमाला पीपीटी, hindi alphabet PPT presentation, hindi varnamala PPT, Hindi Varnamala pdf, हिंदी स्वर, हिंदी व्यंजन, sikhiye hindi varnmala, dr. mulla adam ali, hindi language and literature, hindi alphabet with drawing, hindi alphabet pdf, hindi varnamala for childrens, hindi language, hindi varnamala practice for kids, https://www.drmullaadamali.com
BBR 2024 Summer Sessions Interview Training
Qualitative research interview training by Professor Katrina Pritchard and Dr Helen Williams
Philippine Edukasyong Pantahanan at Pangkabuhayan (EPP) Curriculum
(𝐓𝐋𝐄 𝟏𝟎𝟎) (𝐋𝐞𝐬𝐬𝐨𝐧 𝟏)-𝐏𝐫𝐞𝐥𝐢𝐦𝐬
𝐃𝐢𝐬𝐜𝐮𝐬𝐬 𝐭𝐡𝐞 𝐄𝐏𝐏 𝐂𝐮𝐫𝐫𝐢𝐜𝐮𝐥𝐮𝐦 𝐢𝐧 𝐭𝐡𝐞 𝐏𝐡𝐢𝐥𝐢𝐩𝐩𝐢𝐧𝐞𝐬:
- Understand the goals and objectives of the Edukasyong Pantahanan at Pangkabuhayan (EPP) curriculum, recognizing its importance in fostering practical life skills and values among students. Students will also be able to identify the key components and subjects covered, such as agriculture, home economics, industrial arts, and information and communication technology.
𝐄𝐱𝐩𝐥𝐚𝐢𝐧 𝐭𝐡𝐞 𝐍𝐚𝐭𝐮𝐫𝐞 𝐚𝐧𝐝 𝐒𝐜𝐨𝐩𝐞 𝐨𝐟 𝐚𝐧 𝐄𝐧𝐭𝐫𝐞𝐩𝐫𝐞𝐧𝐞𝐮𝐫:
-Define entrepreneurship, distinguishing it from general business activities by emphasizing its focus on innovation, risk-taking, and value creation. Students will describe the characteristics and traits of successful entrepreneurs, including their roles and responsibilities, and discuss the broader economic and social impacts of entrepreneurial activities on both local and global scales.
Recently uploaded (20)
How to Setup Warehouse & Location in Odoo 17 Inventory
How to Setup Warehouse & Location in Odoo 17 Inventory
BÀI TẬP BỔ TRỢ TIẾNG ANH LỚP 9 CẢ NĂM - GLOBAL SUCCESS - NĂM HỌC 2024-2025 - ...
BÀI TẬP BỔ TRỢ TIẾNG ANH LỚP 9 CẢ NĂM - GLOBAL SUCCESS - NĂM HỌC 2024-2025 - ...
NEWSPAPERS - QUESTION 1 - REVISION POWERPOINT.pptx
NEWSPAPERS - QUESTION 1 - REVISION POWERPOINT.pptx
IGCSE Biology Chapter 14- Reproduction in Plants.pdf
IGCSE Biology Chapter 14- Reproduction in Plants.pdf
Liberal Approach to the Study of Indian Politics.pdf
Liberal Approach to the Study of Indian Politics.pdf
Chapter wise All Notes of First year Basic Civil Engineering.pptx
Chapter wise All Notes of First year Basic Civil Engineering.pptx
spot a liar (Haiqa 146).pptx Technical writhing and presentation skills
spot a liar (Haiqa 146).pptx Technical writhing and presentation skills
Philippine Edukasyong Pantahanan at Pangkabuhayan (EPP) Curriculum
Philippine Edukasyong Pantahanan at Pangkabuhayan (EPP) Curriculum
Subculturing
- 2. Subculture • In biology, a subculture is a new cell or microbiological culture made by transferring some or all cells from a previous culture to fresh growth medium. This action is called sub culturing or passaging the cells. • Subculture is used to prolong the life and/or expand the number of cells or microorganisms in the culture.
- 3. Importance/Role • microorganisms cannot be held in culture for long time due to the gradual rise in toxic metabolites, use of nutrients and increase in cell number due to growth. • Subculture is therefore used to produce a new culture with a lower density of cells than the originating culture, fresh nutrients and no toxic metabolites.
- 4. • Subculture is important for both proliferating (e.g. a microorganism like E. coli) and non- proliferating (e.g. white blood cells) cells. • Typically, subculture is from a culture of a certain volume into fresh growth medium of equal volume, this allows long-term maintenance of the cell line (culture).
- 5. • Subculture into a larger volume of growth medium is used when wanting to increase the number of cells for, for example, use in an industrial process or scientific experiment.
- 6. Culture medium • A growth medium or culture medium is a liquid or gel designed to support the growth of microorganisms or cells, or small plants
- 7. Components of Culture medium • Nutrients: proteins/peptides/amino-acids. • Energy: carbohydrates. • Essential metals and minerals: calcium, magnesium, iron, etc. • Buffering agents: phosphates, acetates etc. • Indicators for pH change: phenol red, bromo-cresol purple etc. • Selective agents: chemicals, antimicrobial agents. • Gelling agent: usually agar. • Vitamins
- 8. Passage number • It is often important to record the approximate number of divisions cells have had in culture by recording the number of passages or subcultures. • In the case of plant tissue cells somaclonal variation may arise over long periods in culture. Similarly in mammalian cell lines chromosomal aberrations have a tendency to increase over time.
- 9. • For microorganisms (contaminations) there is a tendency to adapt to culture conditions, which is rarely quite like the microorganisms in natural environment, which can alter their biology
- 10. Protocols for passaging or sub culturing • The protocol for sub culturing cells depends heavily on the properties of the cells involved. • Non-adherent cells • Adherent cells
- 11. Non-adherent cells • Many cell types, in particular many microorganisms, grow in solution and not attached to a surface. • These cell types can be sub cultured by simply taking a small volume of the parent culture and diluting it in fresh growth medium.
- 12. • Cell density in these cultures is normally measured in cells per milliliter for large eukaryotic cells. • The cells will often have a preferred range of densities for optimal growth and subculture will normally try to keep the cells in this range.
- 13. Adherent cells • Adherent cells, for example many mammalian cell lines, grow attached to a surface such as the bottom of the culture flask. These cell types have to be detached from the surface before they can be sub cultured.
- 15. • For subculture cells may be detached by one of several methods including • Trypsin EDTA (Ethylenediaminetetraacetic Acid) treatment to break down the proteins responsible for surface adherence,
- 16. or • mechanical methods like repeated washing or use of a cell scraper. • The detached cells are then re suspended in fresh growth medium and allowed to settle back onto their growth surface.
- 18. Cell scrapers and cell lifters