( fungicides can be used as a s...)The QIAGEN Guide to Animal Cell Culture

fungicides can be used as a supplement to aseptic technique to prevent microbial contamination. The working concentration of commonly used antibiotics and fungicides is provided in Table 1. Some cell types, particularly primary cells, require additional supplements (e.g., collagen and fibronectin, hormones such as estrogen, and growth factors such as epidermal growth factor and nerve growth factor) to attach to the cell culture vessel and proliferate.

Tip - Media, serum, and supplements should be tested for sterility before use by incubation of a small aliquot at 37C for 48 hours. If microbial growth has occurred after this incubation, the medium or supplement should be discarded.

Tip - The Transfection Tools web site (www.qiagen.com/transfectiontools/) has an online cell database that includes information about media used with different cell types. Click on the cell type of interest, and then click on Detail to view this information.

Table 1. Commonly used antibiotics and fungicides for animal cell culture

Incubation conditions

The incubation conditions used to culture cells are also important. Cell cultures should be incubated in an incubator with a tightly regulated temperature (e.g., a water-jacketed incubator) and CO2 concentration. Most cell lines grow at 37C and 5% CO2 with saturating humidity, but some cell types require incubation at lower temperatures and/or lower CO2 concentrations.

Cell culture vessel

The choice of growth vessel can influence the growth of adherent cells. Sterile, disposable dishes and flasks that have been treated to allow attachme
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