Media Renewal changes two to three times per week.
Rapidly thaw vial by gentle agitation in 37°C water bath (~2 minutes), keeping vial cap out of the water. Decontaminate with 70% ethanol, add 9 mL culture media and centrifuge 125 x g (5-7 minutes). Resuspend in complete culture media and incubate at 37°C in a 5% CO2 atmosphere.
Subculture Ratio: approx. 1.3-1.10
The base medium for this cell line is RPMI, Catalog No. R5886. To make the complete growth medium, add the following components to the base medium: fetal bovine serum, Catalog No. F4135, to a final concentration (v/v) of 10%, L-glutamine, Catalog No. G7513, to a final concentration of 2 mM and sodium pyruvate, Catalog No. S8636, to a final concentration of 1 mM.
The cell freezing medium is DMSO 1X (Catalog Number C6164).