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2. EXPERIMENTAL BACKGROUND
Animal cell culture involves growing the animal cells under controlled conditions in a laboratory culture environment. Cell cultures are obtained through primary culture, the removal of the cells from the organ fragments before or during cultivation, and placed into an appropriate culture environment.
These cells are called primary cells. In summary, primary cells are the ones removed directly from a living tissue and established for growth in vitro.
Hence, primary cells are better resemblance model of functional component of the living tissue as these cells have went through less population doublings compared to continuous cell lines.
When the given culture environment is fully packed with growth of monolayer primary cells (e.g. cell concentration exceeds capacity of given culture environment), growth will stop or greatly reduced.
In this case, subculture must be performed by dividing the culture into
new culture vessels to establish continuous growth of the cell lines.
The following are criteria for subculture:
Cell Culture Basics – A Handbook by Invitrogen and Gibco Freshney, R. I. (2006).
Basic Principles of Cell Culture. Centre for Oncology and Applied Pharmacology.
Experiment in Microbiology and Biochemical Engineering Laboratory Booklet –Department of Biotech. Yonsei University.
Lindl, T. (2002): “Zell- und Gewebekultur.” 5th ed. Spektrum Akademischer Verlag,Heidelberg