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.
Product Format
frozen 1 kit
Storage Conditions
-20°C (or -70°C for long-term storage)
Comments
The Endothelial Cell Growth Kit-VEGF contains components that when added to Vascular Cell Basal Medium (ATCC PCS-100-030) create a complete, low serum culture environment for human large vessel endothelial cells. The final concentration for each component in complete endothelial growth medium is as follows:
rh VEGF: 5 ng/mL
rh EGF: 5 ng/mL
rh FGF basic: 5 ng/mL
rh IGF-1: 15 ng/mL
L-glutamine: 10 mM
Heparin sulfate: 0.75 Units/mL
Hydrocortisone: 1 µg/mL
Ascorbic acid: 50 µg/mL
Fetal bovine serum: 2%
Complete Growth Medium
Obtain one growth kit from the freezer; make sure that the caps of all components 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 Vascular Cell Basal Medium (475 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 media should be stored in the dark at 2°C to 8°C (do not freeze). When stored under these conditions, complete media is stable for 30 days.
Table 1. If using the Endothelial Cell Growth Kit-BBE (ATCC® PCS-100-040), add the indicated volume for each component:
