Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Serum-free medium supports excellent growth curves and normal morphology, as well as serum-free (not animal-free) experimental conditions. The presence of 2% fetal bovine serum in the Fibroblast Growth Kit-Low serum (ATCC PCS-201-041) supports more prolific growth.
Complete Growth Medium
Obtain one growth kit from the freezer; make sure that the caps of all containers are tight.
Thaw the components of the growth kit just prior to adding them to the basal medium. It is necessary to warm the L-glutamine component in a 37°C water bath, and shake to dissolve any precipitates prior to adding to the basal medium.
Obtain one bottle of Fibroblast Basal Medium (480 mL) from cold storage.
Decontaminate the external surfaces of all growth kit component vials and the basal medium bottle by spraying them with 70% ethanol.
Using aseptic technique and working in a laminar flow hood or biosafety cabinet, transfer the volume of each growth kit component, as indicated in Table 1 or 2, to the bottle of basal medium using a separate sterile pipette for each transfer.
Tightly cap the bottle of complete growth medium and swirl the contents gently to assure a homogeneous solution. Do not shake forcefully to avoid foaming. Label and date the bottle.
Complete growth media should be stored in the dark at 2°C to 8°C (do not freeze). When stored under these conditions, complete growth media is stable for 30 days.
Table 1. If using the Fibroblast Growth Kit–Serum-Free (ATCC® PCS-201-040), add the indicated volume for each component in the order shown.