Appendix Cell Culture

In order to study histone mRNA metabolism, we have used MPC-11 mouse myeloma cells that have been adapted to continuous culture, as previously described (58). The mouse myeloma cells were grown in stationary suspension culture in Dulbecco's Modified Eagle's Medium (DMEM) with 10% horse serum, 100 U/ml penicillin, and 100 /xg/ml streptomycin (GIBCO-BRL, Gaithersburg, MD). Mouse myeloma cells are both an economical and a convenient source of material from which both nuclei and polyribosomes can be prepared in quantity. Typically, myeloma cells were grown to a density of 5 X 105 cells/ml and harvested for subcellular fractionation. Up to 38 liters can be grown at once in a large spinner flask (Bellco, Vineland, NJ), but for the most active preparations, 6-8 liters of cells is recommended because of the time required for processing large preparations.

Was this article helpful?

0 0

Post a comment