Introduction to Cell Culture What is Cell Cell culture refers to the removal of
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Introduction to Cell Culture What is Cell Cell culture refers to the removal of cells from an animal or plant and their subsequent The cells may be removed from the tissue may beefrm directly and disaggregated by enzymatic or mechanical means before cultivation, or r cl srain tht has already been already established Primary Culture Primary culture refers to the stage of the culture after the cells are isolated from the tissue and proliferated under the app substrate (i.e, reach confluence). At this stage, the cells have to be subcultured (i.e. passaged) by transferring them to a new vessel with fresh growth medium to provide all of the available te conditions until they occupy a more room for continued growth. Cell Line After the first subculture, the primary culture becomes known as a cell line or subclone. Cell lines derived from primary cultures have a limited life span (ie, they are finite, see below), and as they are passaged, cells with the highest growth capacity predominate, resulting in a degree of genotypic and phenotypic uniformity in the population. Cell Strairn If a subpopulation of a cell line is positively selected from the culture by cloning or some other method, this cell line becomes a cell strain. A cell strain often acquires additional genetic changes subsequent to the initiation of the parent line. Finite vs Continuous Normal cells usually divide only a limited number of times before losing their ability to proliferate, which is a genetically determined event known as senescence; these cell lines are known as finite. However, some cell lines become immortal through a process called transformation, which can occur spontaneously or can be chemically or virally induced When a finite cell line undergoes transformation and acquires the ability to divide indefinitely, it becomes a continuous cell line. Cell Line Culture Conditions Culture conditions vary widely for each cell type, but the artificial environment in which the cells are cultured invariably consists of a suitable vessel containing a substrate or medium that supplies the essential nutrients (amino acids, carbohydrates, vitamins, minerals), growth factors, hormones, and gases (O, CO), and regulates the physico- chemical environment (pH, osmotic pressure, temperature). Most cells are anchorage- dependent and must be cultured while attached to a solid or semi-solid substrate (adherent or monolayer culture), while others can be grown floating in the culture medium (suspension culture) If a surplus of cells are available from subculturing, they should be treated with the appropriate protective agent (e g, DMSO or glycerol) and stored at temperatures below 130 C (cryopreservation) until they are needed. For more information on subculturing and cryopreserving cells, refer to the Guidelines for Maintaining Cultured Cells, page page 26-page page 39 Cryopreservation 2 | Cell Culture Basices é invitrogen gibco LifeExplanation / Answer
1.A). True.
2.A). Sequestered.
3. C). Cells. Proliferation means multiplication.
4.A). True.
5.A). True.
6.B) Two subcultures after the original. Cell lines are produced after the primary culture is passaged several times. So, the first subculture after transferring the primary culture is cell line 1 and next is cell line 2.
7. A).
8. B). Transformation.
9 B). False.
10. False. Some cell lines lose the ability to proliferate but some cells become immortal ( i.e. proliferatee continuously) either spontaneously or when induced chemically or virally.
11 C). Number. Confluence is an estimate of the number of adherent cells in a culture. it is a culture in which all the cells are in contact and the entire surface of the culture medium is covered.
12. B). No.
13. C). Skeletal muscle
14. A). Yes.
15. A). Yes.
16. A). Haemocytometer. It is used to count the number of bacterial cells.
17. A). True.
18. B). False. Most cells are anchorage dependent.
19. A). True. Cell culture is used to test antibiotic sensitivity of bacteria.
20. A). True.
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