Taking Care of Cells in Tissue Culture

Biochemistry, Molecular Biology, and Cell Biology Protocols  >> Taking Care of Cells in Tissue Culture

Here are some general guidelines on maintaining and taking good care of cells in tissue culture:

- split the cells when they are subconfluent; however, it is also not good to split them when they are too dilute; try splitting cells twice a week, in general
- cells generally do not tolerate extreme shifts or changes in temperature, so prewarm media before splitting
- try to be gentle when pipeting in solutions or media for some cells, as it may cause mechanical stress or shearing in fragile cells
- some cells do not tolerate trypsin dissociation so well, so it may be necessary just to use PBS-EDTA to dissociate the cells from the tissue culture plate
- when thawing cells, warm the cells at 37°C as quickly as possible (eg by turning the tube containing the frozen cells in a water bath), preferably within 1-2 minutes, and then adding the thawed cells to fresh media, spinning down the cells, and then replacing with new media, as the DMSO is toxic to the cells
- some cells may initially perform better if incubated in a higher percentage of fetal calf serum (eg 15-20%) instead of the regular 10% serum
- cells generally require time to adapt to change of media, such as when switching to serum-free media, so it may require several passages before they look more healthy; alternatively, it may be prudent to gradually lower the percentage of serum-containing media and increase the percentage of serum-free media with each passage

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