cell line description in detail by krishna

Talking about cell line

1. By Krishna Chavda Unit : 4 - Cell Lines SUBJECT : 401 COURSE : Zoology GIA Roll no. : 4 To : Dr. Divya Chandel

2. Index 1. Cell Culture 2. Cell lines 3. Features of cell lines 4. Characteristics of cell lines 5. Requirements of cell lines 6. Types of cell lines 7. Commonly used cell lines 8. Uses of cell lines 9. Applications of cell lines 10. Limitations of cell lines 11. Questions 12. References

3. Cell Culture : What is cell ? Cells are the basic building blocks of all living things.This complex system is the command for your body. Cell culture : Cell culture is the multifaceted process through which cells are isolated from animal or plant and their subsequent growth under controlled artificial conditions, generally outside their natural environment. In this procedure cells are directly isolated from body or disaggregated by enzymatic or mechanical procedure or they may be derived from cell lines or cell strains. The historical development and methods of cell culture are closely interrelated to those of tissue culture and organ culture. The in vitro propagation of cells has become a common practice in many laboratories for a huge numbers of applications. The ranges of cell types grown are vast.

4. Generally the cells are sensitive to a wide range of compounds and it is therefore necessary to ensure that they come into contact only with those under study and not with extraneous materials. Adherent mammalian cells require a suitable surface for attachment. Primary cell culture : In primary cell culture the cell is first isolated from tissue and then proliferated under the appropriate conditions until they consume all available contents for their growth. Now the cell is ready for subculturing by transferring them to new growth medium that furnish more opportunity for continued growth. Type of cell culture : Continuous cell culture A continuous cell culture is one that is apparently capable of an unlimited number of population doublings, often referred to as an immortal cell culture. 2. Finite cell culture A finite cell culture is capable of only a limited number of population doublings after which the culture ceases proliferation.

5. Stages of cell culture : Isolated tissue Disaggregation Primary cell culture Finite cell cultures Continuous cell lines

6. Cell line : A cell line is a permanently established cell culture that will proliferate indefinitely in appropriate fresh medium and space. In January, 1912, Carrel and his coworkers developed the first “cell line” derived from the fragments of explanted chicken embryo heart. This cell line was subcultured hundreds of times, and after the initial contamination outbreak, it was continued by Arthur Ebeling in Carrel's laboratory. History of cell line :

7. To use any cell line for the production of biological product, one should have knowledge of following things related to cell lines: • Age, sex and species of the donor tissue. • For human cell lines, the donor’s medical history and if available, the results of tests performed on the donor for the detection of adventitious agents. • Culture history of the cell line including methods used for the isolation of the tissues from which the line was derived, passage history, media used and history of passage in animals, etc. • Previous identity testing and the results of all available adventitious agents testing. Features of cell lines :

8. Characteristics of cell lines : Each cell line is distinguished by characteristic features which render these cells unique and biomedically or biotechnologically useful. • The growth pattern and morphological appearance of the cell line should be determined and should be stable from the master cell bank to the end-of-production cells. • If there are specific markers that may be useful in characterizing the cell line (such as marker chromosomes, specific surface markers), these should be characterized for stability. • Mostly cultured cell lines are allowed to generate their own ECM (extra cellular matrix), but primary culture and propagation of some specialized cells, exogenous provision of ECM.

9. • Many transformed cell lines have provided the best model for the induction of differentiation. • Since normal cells has limited dividing capacity, therefore after a fixed number of population doublings cell lines derived from normal tissue will die out. This is a genetically determined event involving several different genes and this phenomenon is known as senescence. If the cells have an identified finite life expectancy, the total number of population doubling levels through senescence should be determined. • Some cell lines may avoid senescence and give rise to continuous cell lines. The ability of a cell line to grow continuously probably reflects its capacity for genetic variation, allowing subsequent selection. • A common feature of many human continuous cell lines is the development of a subtetraploid chromosome number. The alteration in a culture that gives rise to a continuous cell line is commonly called in vitro transformation and may occur spontaneously or be chemically or virally induced.

10. For the maintenance of Cell line some basic conditions are required. These are described as follows. 1. pH: Most cell lines grow well at pH 7.4. Although the optimum pH for cell growth varies relatively little among different cell strains, some normal fibroblast lines perform best at pH 7.4 to pH 7.7, and transformed cells may do better at pH 7.0 to pH 7.4. 2. Buffering: Culture media must be buffered under two sets of conditions: a) Open dishes, where the evolution of CO2 causes the pH to rise b) Overproduction of CO2 and lactic acid in transformed cell lines at high cell concentrations, when the pH will fall. 3. Temperature: The temperature recommended for most human and warm-blooded animal cell lines is 37°C, closely to body heat, but generally set a little lower for safety, because overheating may become major problem than under heating. Requirements of cell lines :

11. 4. Media: Although many cell lines are still propagated in medium supplemented with serum, in many instances cultures may now be propagated in serum-free media. Media that have been produced commercially will have been tested for their capability of sustaining the growth of one or more cell lines. 5. Growth curve: A growth curve gives three parameters of measurement: (1) the lag phase before cell proliferation is initiated after subculture, indicating whether the cells are having to adapt to different conditions; (2) the doubling time in the middle of the exponential growth phase, indicating the growth promoting capacity of the medium; and (3) the maximum cell concentration attainable indicating whether there are limiting concentrations of certain nutrients. In cell lines whose growth is not sensitive to density (e.g., continuous cell lines), the terminal cell density indicates the total yield possible and usually reflects the total amino acid or glucose concentration.

12. Fibroblast Types of cell line Cell lines Morphology Origin Species Age Ploidy Characteristics IMR-90 MRC-5 Fibroblast Fibroblast Lung Human Embryonic Diploid Lung Human Embryonic Diploid Susceptible to human viral infection contact inhibited Finite from normal tissue Continuous from normal tissue HaCaT Epithelial Keratinocytes Human Adult Diploid Comification BRL3A Epithelial Liver Rat New born Produce IGF-2 - Continuous from neuroplastic tissue HeLa Epithelial Cervix Human Adult Aneuploid G6PD Type A B16 Fibroblast Melanoma Mouse Adult Aneuploid Melanin Types of cell lines :

13. Commonly used cell lines : Several cell lines are widely used in contemporary research across various fields. Some of the popularly used cell lines : 1. HeLa Cells : Derived from cervical cancer cells, HeLa cells are one of the most famous and commonly used cell lines in biomedical research. They have been instrumental in various areas, including cancer research and virology. 2. HEK 293 Cells : These are human embryonic kidney cells that are widely used for the production of recombinant proteins and the study of gene expression and protein function. 3. A549 Cells : Derived from human lung adenocarcinoma, A549 cells are used in lung cancer research, drug testing, and studies related to respiratory diseases. HeLa Cells HEK 293 Cells A549 Cells

14. 4. MCF-7 Cells : These are breast cancer cells used for breast cancer research and studies on hormone receptor-positive breast cancer. 5. SH-SY5Y Cells : Derived from human neuroblastoma, these cells are used in neuroscience research and drug discovery related to neurodegenerative diseases. 6. U-87 MG Cells : A glioblastoma cell line often used in brain cancer research and studies on central nervous system disorders. MCF-7 Cells SH-SY5Y Cells U-87 MG Cells

15. Uses of cell lines : Cell lines are in vitro model systems that are widely used in different fields of medical research, especially basic cancer research and drug discovery. Their usefulness is primarily linked to their ability to provide an indefinite source of biological material for experimental purposes. Being used in vaccine production, testing drug metabolism and cytotoxicity, antibody production, study of gene function, generation of artificial tissues (e.g., artificial skin) and synthesis of biological compounds e.g., therapeutic proteins.

16. Applications of cell lines : 1. Biomedical Research : Cell lines are fundamental tools for studying basic cellular processes such as cell division, differentiation, and apoptosis. They serve as models for understanding disease mechanisms, especially genetic disorders. 2. Drug Screening and Development : Cell lines are used in high-throughput screening to test the efficacy and toxicity of potential drugs, accelerating drug discovery. 3. Vaccine Development : Viral cell lines are employed to produce viral proteins used in vaccine development, like the influenza vaccine. 4. Cancer Research : Cancer cell lines help researchers investigate the molecular basis of cancer, screen for potential anticancer compounds, and study drug resistance.

17. 5. Toxicology Testing : Cell lines are used to assess the toxicity of chemicals, drugs, and environmental agents, reducing reliance on animal models. 6. Gene Expression Studies : Researchers study gene regulation, expression patterns, and the effects of genetic mutations using cell lines. 7. Bioproduction : Some cell lines, such as Chinese hamster ovary (CHO) cells, are used in biotechnology to produce biopharmaceuticals like monoclonal antibodies and insulin.

18. Limitations of cell lines : 1. Genetic Stability : Cell lines can accumulate genetic changes over time, leading to differences from the original tissue. This instability can affect experimental reproducibility. 2. Lack of Tissue Complexity : Cell lines lack the three-dimensional structure, cell-cell interactions, and physiological conditions found in intact tissues and organisms. 3. Contamination : Cross-contamination or misidentification of cell lines can result in unreliable data. Authentication is crucial to ensure the cell line's identity. 4. Limited Lifespan : Most cell lines have a finite lifespan and eventually enter a state of senescence, limiting their use for long-term studies.

19. 5. Phenotypic Changes : Cell lines can exhibit different characteristics from the original tissue due to adaptation to in vitro conditions. This can lead to discrepancies between cell line behavior and in vivo biology. 6. Ethical Concerns : Human cell lines, especially those derived from questionable sources, raise ethical concerns regarding informed consent, privacy, and usage. 7. Relevance to In Vivo Conditions : Findings from cell line studies may not always translate directly to in vivo situations, necessitating animal or clinical studies. In conclusion, cell lines are valuable tools in scientific research, but researchers must be aware of their limitations. Careful experimental design, regular authentication, and complementing cell line studies with in vivo research are essential for maximizing the reliability and relevance of the findings obtained from cell lines.

20. Questions : 7 marks : 1. Describe cell lines in detail ? 2. Give the applications and limitations of cell lines.

21. 1 marks : 1. Define : Cell Culture Cell culture is the multifaceted process through which cells are isolated from animal or plant and their subsequent growth under controlled artificial conditions, generally outside their natural environment. 2. Give any two features of cell lines. For human cell lines, the donor’s medical history and if available, the results of tests performed on the donor for the detection of adventitious agents. Culture history of the cell line including methods used for the isolation of the tissues from which the line was derived, passage history, media used and history of passage in animals, etc.

22. 3. Which are the main three types of cell lines ? Finite from normal tissue Continuous from normal tissue Continuous from neuroplastic tisuue 4. Give any three name of the commonly used cell lines. HeLa HEK 293 MCF-7 5 .Describe in short : Uses of cell lines. Cell lines are being used in vaccine production, testing drug metabolism and cytotoxicity, antibody production, study of gene function, generation of artificial tissues (e.g., artificial skin) and synthesis of biological compounds e.g., therapeutic proteins.

23. MCQs: 1. Which is finite cell line from given options ? a) HeLa b) MCF-7 c) MRC-5 d) HEK 293 2. Comification is characteristics of which cell line ? a) IMR-90 b) HaCat c) B16 d) MRC-5 3. Who is the develop cell lines for first time ? a) Arthur Ebeling b) Herbert Boyer c) Robert Hooke d) Carrel 4. Which cells are use in Vaccine production ? a) Cancer cells b) Chinese hamster ovary c) Viral cells d) U-87 MG Cells 5. BRL3A cell lines are found in which species ? a) Mouse b) Rat c) Human d) Pig √ √ √ √ √

24. References : https://www.mlsu.ac.in/econtents/315_Unit%203-%20Nomenclature%20and%20Maintenance.pdf https://www.researchgate.net/publication/333042040_CELL_LINE_A_REVIEW https://www.slideshare.net/Vijayanarasimha1/cell-lines

25. Thank you