I'm currently using RPMI 1640 (plus 10% FBS, 55 μM 2-mercaptoethanol, 1mM sodium pyruvate,1mM HEPES, 1mM NEAA,1% antibiotic-antimyetic ) for BM-DCs and MLR. Any suggestions to improve this protocol?
Thanks~!
For the MLR: RPMI 1640 plus 5% FBS + 50 μM 2-mercaptoethanol + 20 mM L-glutamine will be sufficient. You can use also 5% heat-inactivated human serum instead of FBS. In long term cultures the concentration of serum should be increased to 10%. HEPES is required only if many operations are performed outside CO2-incubator and significantly increase pH. Accurate handling allows not to use antibiotics, but you can add them into primary cultures. I successfully grew T cell clones without antibiotics, sodium piruvate and NEAA. I have no experience with BM-DC. It may be another scientist can help.
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