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Chapter 10.3 : Cell Culture Techniques: Serial Propagation and Maintenance of Monolayer Cell Cultures

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Cell Culture Techniques: Serial Propagation and Maintenance of Monolayer Cell Cultures, Page 1 of 2

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Abstract:

The majority of diagnostic laboratories that perform virus culture obtain the requisite cells from a commercial vendor, already seeded in the vessels that will be inoculated. Alternatively, they receive them in a near-ready state, with minimal manipulations required before the cultures are ready to inoculate. Few laboratories still maintain in-house seed stocks and routinely culture, passage, and prepare cultures for virus culture work. For those that choose to, it is imperative that they have the appropriate facilities and adhere strictly to detailed, well-established protocols, to ensure an uninterrupted supply of healthy cell cultures for the diagnostic laboratory. The procedure detailed below provides information on these procedures and is revised from the previously published edition ( ).

Citation: Leber A. 2016. Cell Culture Techniques: Serial Propagation and Maintenance of Monolayer Cell Cultures, p 10.3.1-10.3.10. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.3
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References

/content/book/10.1128/9781555818814.chap10.3
1. Hodinka RL. 2010. Cell culture techniques: serial propagation and maintenance of monolayer cell cultures. In Garcia LS (ed), Clinical Microbiology Procedures Handbook, 3rd ed. ASM Press, Washington, DC.
2. Hayflick L. 1965. The limited in vitro lifetime of human diploid cell strains. Exp Cell Res 37:614636.
3. Freshney RI. 1983. Culture of Animal Cells: a Manual of Basic Technique. Alan R. Liss, Inc., New York, NY.
4. Bird BR, Forrester FT. 1981. Basic Laboratory Techniques in Cell Culture. U.S. Department of Health and Human Services, CDC, Atlanta, GA.
1. Bird BR, Forrester FT. 1981. Basic Laboratory Techniques in Cell Culture. U.S. Department of Health and Human Services, CDC, Atlanta, GA.
2. McGarrity GJ. 1982. Detection of mycoplasma infection of cell cultures, p 99131. In Maramorosch K (ed), Advances in Cell Culture. Academic Press, Inc., New York, NY.
3. Schmidt NJ. 1989. Cell culture procedures for diagnostic virology, p 51100. In Schmidt NJ, Emmons RW (ed), Diagnostic Procedures for Viral, Rickettsial, and Chlamydial Infections, 6th ed. American Public Health Association, Washington, DC.
4. van Kuppeveld FJ, Johansson KE, Galama JM, Kissing J, Bölske G, van der Logt JT, Melchers WJ. 1994. Detection of mycoplasma contamination in cell cultures by a mycoplasma group-specific PCR. Appl Environ Microbiol 60:149152.
1. Bird BR, Forrester FT. 1981. Basic Laboratory Techniques in Cell Culture. U.S. Department of Health and Human Services, CDC, Atlanta, GA.

Tables

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Table 10.3–1

Characteristics of monolayer cell cultures

Citation: Leber A. 2016. Cell Culture Techniques: Serial Propagation and Maintenance of Monolayer Cell Cultures, p 10.3.1-10.3.10. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.3
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Table 10.3–2

Reagent volumes for trypsinization of monolayer cell cultures

Citation: Leber A. 2016. Cell Culture Techniques: Serial Propagation and Maintenance of Monolayer Cell Cultures, p 10.3.1-10.3.10. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.3
Generic image for table
Table 10.3–3

Recommended split ratios for commonly used cell lines

Citation: Leber A. 2016. Cell Culture Techniques: Serial Propagation and Maintenance of Monolayer Cell Cultures, p 10.3.1-10.3.10. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.3
Generic image for table
Table 10.3–4

Surface area, cell yield, and final volume of growth medium for commonly used culture vessels

Citation: Leber A. 2016. Cell Culture Techniques: Serial Propagation and Maintenance of Monolayer Cell Cultures, p 10.3.1-10.3.10. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.3

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