A cell line is a product of immortal cells that are used for biological research.
Cells used for cell lines are immortal, that happens if a cell is cancerous.
The cells can perpetuate division indefinitely which is unlike regular cells which can only divide approximately 50 times.
Human cell lines
MCF-7 breast cancer
HL 60 Leukemia
HEK-293 Human embryonic kidney
HeLa Henrietta lacks
Primate cell lines
Vero African green monkey kidney epithelial cells
Cos-7 African green monkey kidney cells
And others such as CHO from hamster, sf9 & sf21 from insect cells.
Biology and characterization of the cell cultureKAUSHAL SAHU
Introduction
History
Important terminology
Biology of culture cell
Characterization of culture cell
Application of animal culture
Conclusion
References
Introduction
Definition
History
Why are the transgenic animals being produced
Transgenic mice
Mice: as model organism
Methods of creation of transgenic mice
knock-out mice
Application of transgenic mice
Conclusion
References
Equipments used , types of culture and media, subculturing, secondary culture, finite & continuous cell lines, cryopreservation and applications of cell culture
Introduction.
Properties of Stem Cells.
Key Research events.
Embryonic Stem Cell.
Stem cell Cultivation.
Stem cells are central to three processes in an organism.
Research & Clinical Application of stem cell.
Research patents.
Conclusion.
Reference.
In vitro methods for the assessment of general cellular toxicity,
End-points for the assessment of general cellular toxicity
Specialized cells commonly used in toxicology
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
Biology and characterization of the cell cultureKAUSHAL SAHU
Introduction
History
Important terminology
Biology of culture cell
Characterization of culture cell
Application of animal culture
Conclusion
References
Introduction
Definition
History
Why are the transgenic animals being produced
Transgenic mice
Mice: as model organism
Methods of creation of transgenic mice
knock-out mice
Application of transgenic mice
Conclusion
References
Equipments used , types of culture and media, subculturing, secondary culture, finite & continuous cell lines, cryopreservation and applications of cell culture
Introduction.
Properties of Stem Cells.
Key Research events.
Embryonic Stem Cell.
Stem cell Cultivation.
Stem cells are central to three processes in an organism.
Research & Clinical Application of stem cell.
Research patents.
Conclusion.
Reference.
In vitro methods for the assessment of general cellular toxicity,
End-points for the assessment of general cellular toxicity
Specialized cells commonly used in toxicology
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
Animal cell culture in Biopharmaceutical Industry in the Production of Therap...Shubham Chinchulkar
This presentation will help you to understand the basics of Animal cell culture along with its applicability in the diagnosis and treatment of cancer, and autoimmune diseases.
This presentation will help to understand the basics of mammalian cell culture. I have also covered the difference between adherent and suspension cell lines. I have also included the advantages and disadvantages of the cell line.
SYNTHETIC CELLS
An artificial cell or minimal cell or synthetic cell is an engineered particle that mimics one or many functions of a biological cell.
Artificial cells are biological or polymeric membranes which enclose biologically active materials.
A "living" artificial cell has been defined as a completely synthetically made cell that can capture energy, maintain ion gradients, contain macromolecules as well as store information and have the ability to mutate.
DEFINITION
EXAMPLE
SYNTHETIC BIOLOGY
Synthetic biology is a multidisciplinary area of research that seeks to create new biological parts, devices, and systems, or to redesign systems that are already found in nature.
Due to more powerful genetic engineering capabilities and decreased DNA synthesis and sequencing costs, the field of synthetic biology is rapidly growing
HISTORY
BOTTOM-UP APPROACH FOR CONSTRUCTING SYNTHETIC CELLS
A bottom-up approach is commonly used to design and construct genetic circuits by piecing together functional modules that are capable of reprogramming cells with novel behavior.
CELL ENCAPSULATION METHOD
Cell microencapsulation technology involves immobilization of the cells within a polymeric semi-permeable membrane that permits the bidirectional diffusion of molecules such as the influx of oxygen, nutrients, growth factors etc. essential for cell metabolism and the outward diffusion of waste products and therapeutic proteins.
TECHNIQUES USED FOR THE PREPARATION OF EMULSION
1- high pressure homogenization
2- microfluidization
3- drop method
4- emulsion method
MEMBRANES OF SYNTHETIC CELLS
THE MINIMAL CELL
A minimal cell is one whose genome only encodes the minimal set of genes necessary for the cell to survive.
THE SYNTHETIC BLOOD CELLS
Synthetic red blood cells mimic natural ones, and have new abilities
APPLICATIONS OF SYNTHETIC CELLS
1- DRUG RELEASE AND DELIEVERY
2- GENE THERAPY
3- ENZYME THERAPY
4- HEMOPERFUSION
5- OTHER APPLICATIONS
FUTURE OF SYNTHETIC CELLS AND BIOLOGY
ACHIEVEMENTS
HEALTH AND SAFETY ISSUES
ETHICS AND CONTROVERSIES
REFERENCES
THANK YOU
Different applications of Animal cell culture:
Model Systems
Toxicity Testing
Drug Screening and Development
Virology
Genetic Engineering
Gene Therapy
Stem Cell Therapy
Disease Diagnosis
Cancer Research
Cell-based Manufacturing
Production of vaccines
Recombinant proteins
Production of Biopesticides
This slide explains the various basic aspect of animal cell culture, cell line and cell strain, initiation and maintenance of primary cell culture, characteristic of primary cell culture and their applications. It also contains MCQs for practice.
Reprogramming to pluripotency is possible from adult cells of different tissues and species through the ectopic expression of defined factors. The generated induced Pluripotent Stem Cells (iPSCs) are relevant for various purposes, including disease modeling, drug or toxicity screening and autologous cell therapy. Over the last few years, increased efforts are being made to improve the reprogramming techniques, the efficiency and quality of the generated iPSCs, as well as to identify the best cell source to be reprogrammed. Cells derived from fetal tissues, such as amniotic fluid, placenta and umbilical cord, offer distinct advantages in terms of reprogramming compared to adult somatic cells. Importantly, fetal cells are more primitive, easily achievable in sufficient numbers and are devoid of any ethical concern. They show great plasticity, high proliferation rate, low immunogenity and absence of teratoma formation. Therefore, they can be reprogrammed much faster and more efficiently than adult cells. Here, we provide a comprehensive overview of the advantages of reprogramming fetal sources in comparison to other commonly used cell types.
Animal Cell culture by S.D.Mankar, Pravara Rural college of Pharmacysomeshwar mankar
Growth of animal cells in culture, general procedure for cell culture,
Primary, established and transformed cell cultures.
Application of cell cultures in pharmaceutical industry and research.
Viruses are obligate intracellular parasites which means they can only grow or reproduce inside a host cell.
The primary purpose of virus cultivation:
To isolate and identify viruses in clinical samples.
To do research on the viral structure, replication, genetics, and effects on the host cell.
To prepare viruses for vaccine production.
Isolation of the virus is always considered a gold standard for establishing the viral origin of the disease
topics covered
CULTIVATION OF VIRUSES
Animal inoculation
Embryonated eggs
CAM
Allantoic cavity
Amniotic cavity
Yolk sac
Tissue culture
Organ culture
Explant culture
Cell culture
Primary cell culture
diploid cell culture
Continues cell lines
Introduction
History
Tumor suppressor gene- pRB
- RB gene
- Role of RB in regulation of cell cycle
- Tumor associated with RB gene mutation
Tumor suppressor gene- p53
- What is p53 gene?
- Function of p53 gene
- How it regulates cell cycle
- What happen if p53 gene inactivated
- Cancer associated with p53 mutation
- Conclusion
- References
Introduction
Definition
History
Two hit hypothesis
Functions
Mutation in tumor suppressor genes
What is mutation
Inherited mutation of TSGs
Acquired mutation of TSGs
What is Oncogenes?
TSGs and Oncogenes : Brakes and accelerators
Stop and go signal
Examples of TSGs:
RB-The retinoblastoma gene
P53 protein
TSGs &cell suicide
Conclusion
References
Introduction
Protein synthesis
Synthesis of secretory proteins on membrane-bound ribosomes
Processing of newly synthesized proteins in the ER
Synthesis of integral membrane protein on membrane bound ribosomes
Maintenance of membrane asymmetry
Conclusion
Reference
Introduction
Definition
Factors required for Translation
Formation of aminoacyl t-RNA
1)Activation of amino acid
2) Transfer of amino acid to t-RNA
Translation involves following steps:-
1)Initiation
2)Elongation
3)Termination
Conclusion
Reference
Introduction
Definition
History
central dogma
Major components
mRNA,tRNA,rRNA
Energy source
Amino acids
Protien factor
Enzymes
Inorganic ions
Step involves in translation:
Aminoacylation of tRNA
Initiation
Elongation
termination
Importance of translation
Conclusion
Reference
Introduction
Protein modifications
Folding
Chaperon mediated
Enzymatic
Cleavage
Addition of functional groups
Chemical groups
Hydrophobic groups
Proteolysis
Conclusion
Reference
INTRODUCTION
HISTORY
WHAT IS TRANSCRIPTION
PROKARYOTIC TRANSCRIPTION
STEPS OF TRANSCRIPTION
HOW TRANSCRIPTION OCCURS
PROCESS OF TRANSCRIPTION
Initiation
Elongation
Termination
CONCLUSION
REFRENCES
Enzyme Kinetics and thermodynamic analysisKAUSHAL SAHU
Introduction
Kinetics and thermodynamicSG
Thermodynamic in enzymatic reactions
balanced equations in chemical reactions
changes in free energy determine the direction & equilibrium state of chemical reactions
the rates of reactions
Factors effecting enzymatic activity
(i) Enzyme concentration.
(ii) Substrate concentration.
(iii)Temperature
(iv) pH.
(v) Activators.
(vi)Inhibitors
Michaelis-menten equation
CONCLUSIONS
REFERENECES
Recepter mediated endocytosis by kk ashuKAUSHAL SAHU
INTRODUCTION
DEFINITION OF RECEPTOR MEDIATED ENDOCYTOSIS
WHAT TYPE OF LIGANDS ENTER BY RME?
FORMATION OF CLATHRIN-COATED VESICLES
TRISKELIONS
ROLE OF DYNAMIN IN THE FORMATION OF CLATHRIN-COATED VESICLES
ROLE OF PHOSPHOLIPIDS IN THE FORMATION OF COATED VESICLES
ENDOCYTIC PATHWAY
LDLs AND CHOLESTROL METABOLISM
CONCLUSION
REFERENCES
The delivery of newly synthesized protein to their proper cellular destination, usually referred to as protein targeting or sorting.
The mode of protein transport depends chiefly on the location in the cell cytoplasm of the polysomes involved in protein synthesis.
There are two modes of protein sorting:-
1) Co - translational Transportation.
2) Post - translational Transportation.
Prokaryotic translation machinery by kk KAUSHAL SAHU
Introduction
Definition
Factors required for Translation
Formation of aminoacyl t-RNA
1)Activation of amino acid
2) Transfer of amino acid to t-RNA
Translation involves following steps:-
1)Initiation
2)Elongation
3)Termination
Conclusion
Reference
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
In silico drugs analogue design: novobiocin analogues.pptx
Applications of cell lines
1. Applications of
cell lines
By
KAUSHAL KUMAR SAHU
Assistant Professor (Ad Hoc)
Department of Biotechnology
Govt. Digvijay Autonomous P. G. College
Raj-Nandgaon ( C. G. )
2. Cell culture
Cell culture is the process by which prokaryotic, eukaryotic or plant cells are grown
under controlled conditions. But in practice it refers to the culturing of cells derived
from animal cells.
Cell culture was first successfully undertaken by Ross Harrison in 1907
Roux in 1885 for the first time maintained embryonic chick cells in a cell culture.
3. Why is cell culture used for?
Areas where cell culture technology is currently playing a major role.
Model systems for
Studying basic cell biology, interactions between disease causing agents and cells,
effects of drugs on cells, process and triggering of aging & nutritional studies
Toxicity testing
Study the effects of new drugs
Cancer research
Study the function of various chemicals, virus & radiation to convert normal cultured
cells to cancerous cells
4. Conti…..
Virology
Cultivation of virus for vaccine production, also used to study there infectious cycle. e.g.
polio, rabies, chicken pox, hepatitis B & measles.
Gene therapy
Cells having a functional gene can be replaced to cells which are having non-functional
gene.
5. Tissue culture
•In vitro cultivation of organs, tissues & cells at defined temperature using an incubator
& supplemented with a medium containing cell nutrients & growth factors is collectively
known as tissue culture.
•Different types of cell grown in culture includes connective tissue elements such as
fibroblasts, skeletal tissue, cardiac, epithelial tissue (liver, breast, skin, kidney) and many
different types of tumor cells.
6. Primary culture
•Cells when surgically or enzymatically removed from an organism and placed in suitable
culture environment will attach and grow are called as primary culture.
•Primary cells have a finite life span.
•Primary culture contains a very heterogeneous population of cells.
•Treated by proteolytic enzyme (Trypsin).
7. Cell lines
A cell line is a product of immortal cells that are used for biological research.
Cells used for cell lines are immortal, that happens if a cell is cancerous.
The cells can perpetuate division indefinitely which is unlike regular cells which can only
divide approximately 50 times.
WRL-68HeLa Jurkat
8. Common cell lines
Human cell lines
MCF-7 breast cancer
HL 60 Leukemia
HEK-293 Human embryonic kidney
HeLa Henrietta lacks
Primate cell lines
Vero African green monkey kidney epithelial cells
Cos-7 African green monkey kidney cells
And others such as CHO from hamster, sf9 & sf21 from insect cells.
10. Continuous cell lines
Cell lines which either occur spontaneously or induced virally or chemically
transformed into Continuous cell lines
Characteristics of continuous cell lines
• -smaller, more rounded, less adherent with a higher nucleus /cytoplasm
ratio
• -Fast growth and have aneuploid chromosome number
14. What is in the media?
Dulbecco’ Modified Eagle’s Media (DMEM)
Contains glucose, some proteins, and essential salts
Contains a pH indicator (phenol red) Media looks pink/red at pH 7.2
Acidic -yellow or orange (cell growth, bacterial growth)
Basic -purple (no cell growth, not enough CO2)
Antibiotics (penicillin and streptomycin)
Prevent bacterial contamination
Salts and buffers
To simulate in vivo environment
15. Conti……
Serum
Portion of blood after the cells and fibers have clotted
From cow, horse, sheep
added to media as a nutrient source for growing cells
Lipids, proteins
16. Use of tissue cultures in toxicity
testing
Mammalian cell cultures can be a suitable alternative for the use of whole animal tests
to establish the potential toxicity of compounds.
This due to many reasons:
•They can overcome the disadvantages of the whole animal tests including:
High cost
Variability of results
•Cell culture tests are ripid,allow more efficient screening of novel compounds and sometimes can
allow identification of metabolic target of inhibition.
17. Conti…..
Cell culture tests can be designed to evaluate various effect:
Reduced growth rate
Breakdown of membrane permeability
Tissue specificity response
Ability to metabolize toxic compound
Genetic effect
18. Use of tissue culture for biological
products
Production of vaccines:
• Two factors stimulated the use of tissue cultures for vaccine production:
▫ The ability to grow viruses in cell culture
Current egg vaccine production require long time(9 month) that hinder the response to
unanticipated demands.
In (1949),Enders discovered that the poliomyelitis virus could be grown from primary
monkey cells in culture.
The polio vaccine, produced in 1954, was the first human vaccine to be produced using
large-scale cell culture techniques.
19. Conti…
Animal cell technology is considerably developed for the production of
a range of human and veterinary viral vaccines against a variety of
diseases.
(b) Production of antibodies:
Also, the in vitro methods for production of mABs are the methods of
choice because of:
The ease of culture for production.
Less economic consideration compared with the use of animals.
20. Conti…..
Practical uses of the in vitro produced mABs:
Diagnostic tests for the identification of small quantities of specific antigens.
mABs also are used therapeutically: OKT3 recognizes a surface antigen (CD3) on T cell
and is one of the most effective agents in preventing immunological rejection of
transplanted kidneys.
21. Conti…..
Various mAbs designed to destruct tumor cells by targeting a membrane bound
protein antigens specifically expressed by these cells.
The conjugation of radiactive or toxic compounds to the antibody can result in a
localized high concentration resulting in cytotoxicity to the target cells.
23. Conti….
Some examples for these biological products:
Interferone:
Discovered when Isaacs and Lindenmann (1957) found that culture medium taken from cells that
had supported viral growth could protect non-infected cells from a subsequent viral infection.
Tissue plasminogen activator (t-PA ):
t-PA was produced in large scale by Genenteck from transfected CHO-K1 cells. It is used to prevent
undesirable formation fibrin clots in the bloodstream.
Blood clotting factors:
For example, factor VIII is produced in large scale by Bayer through transfection of the mammalian
kidney cell line (BHK) with an appropriate gene.
24. Cell therapy
Literally, cell therapy means treatment with cells, i.e replacing diseased or
dysfunctional cells with healthy functioning ones.
For example:
•When hematopoietic cells are vulnerable to destruction by any cytotoxic drugs used in
chemotherapy to eradicate residual tumor cells.
25. Drug screening & development
• Cell based assay 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 of drugs. Originally these cell culture tests were done in 96
well plates, but increasing use is now being made of 384 & 1536 well plate.
Corning micro plate
26. Respiratory cell line
Alveolar cells
A549 cells
Bronchial epithelial cells
BEAS 2B, DMS53, SHP-77,
NCI-H23, 292, 524, 727,11655,1299
HBE4-E6/E7 etc….. (>140)
Macrophage
Differentiated U937/MonoMac6 cell
Murine RAW264.7
• Respiratory cell lines are useful for evaluating of molecular
mechanisms of inflammation and other physiological /pathological
events.
27. Pancreatic beta cell lines and their applications
in diabetes mellitus research
During the past 30 years great effort has been put into establishing an insulin-
secreting beta cell line that retains normal regulation of insulin secretion, but only few
of these attempts have been successful.
To overcome the limited availability of primary beta cells into the field of diabetes
mellitus research, numerous investigators used X-rays or viruses to induce insulinomas,
in vitro transformation, derivation of cells from transgenic mice or even non-islet cells to
produce immortalized beta cell lines.
28. The most widely used insulin-secreting cell lines are RIN, HIT, MIN, INS-1 and TC
cells.
These cells produce insulin and small amounts of glucagon and somatostatin.
Despite problems associated with beta cell cultures, these cell lines have provided
some valuable information about physiological processes.
However, an urgent need to establish a "normal" beta cell line of human or pig
origin remains.
29. Vivalis, a Nantes, France-based biotechnology company, has developed EB66®, a
novel cell line derived from duck embryonic stem cells for the cell culture
production of viral vaccines.
As a result, EB66 cells have become a superior industry alternative for the safe,
cost effective manufacturing of viral vaccines.
Cell line in vaccinesCell lines in vaccines
30. Vivalis offers research and commercial licenses for this cell line to pharmaceutical
and biotechnology companies for the production of prophylactic and therapeutic
vaccines.