DT40 cell culture media recipe
DT40s are a cell culture line of chicken lymphocyte B cells. They exhibit a high rate of targeted (and random) integration of transfected DNA constructs and are ideal for stable transfection. They can, however, be a royal pain to culture. Anyone who has worked with DT40s can attest to the fact that it is probably the most finicky cell line they have ever worked with. The key to keeping the DT40s alive and thriving is knowing how often to split them (i.e. not letting them get too dense while not splitting them too early…80% plate density is just about right) and knowing what environment (i.e. cell culture medium) to grow them in. Through trial and error, a good friend of mine used the following culture media recipe for both her wild-type and stably transfected DT40s (for experiments that eventually got published in Nature Cell Biology)
To make 1 Liter:
RPMI 1640 Medium (w/ sodium bicarb & w/o glutamine) - 875 mL
1 microMolar glutamine - 5 mL
10% fetal bovine serum - 100 mL
1% chicken serum - 10 mL
50 microMolar 2-mercaptoethanol - 3.5 microliters
Penn/Strep (0.5 mg/mL G418) - 10 mL
Sterile filter.
