PowerPoint Contents
Introduction
Mammalian cell plating efficiency involves cultivating cells in vitro at low cell density until formation of cell colonies. Plating efficiency can be used to determine the optimal culture media, culture supplements, and environmental conditions necessary for cell growth, as determined by a high plating efficiency. Note the two discrete Chinese hamster ovary cell colonies in the upper left and lower right of the figure, and the clump of unattached dead cells in the lower left corner. The cells were visualized using an inverted light microscope with a 40X objective (Colony 12083).
Methods
Chinese hamster ovary cells were treated with trypsin to detach cells and make a single cell suspension, followed by cell enumeration using a hemacytometer. Cells were diluted to a concentration of 1,000 cells/ml, serially diluted in various culture media, and plated into 96-well plates. The plates were incubated for 10 days at 37°C and 5% CO2in Ham's F-12 medium containing 5% fetal calf serum. The plating efficiency is calculated for a set of culture conditions using the formula: Number of colonies formed/Number of cells seeded X 100 = Plating efficiency (%).
Discussion
Mammalian cells grown in culture are required for the multiple research and applied applications in bioscience. In microbiology, cell cultures are used for propagation of viruses for isolation, identification, and antiviral drug testing. Determining optimal growth conditions for host cells is a crucial first step in optimizing conditions for replication of obligate intracellular pathogens. Plating efficiency is a method used to optimize growth conditions.
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Slide 4: Labeled View
Contact Information
ASM Education, education@asmusa.org