Recently, I am planing to use 3mM free fatty acid to treat the HepG2 cells. I am looking for the protocol for this.
My protocol:
Working solution preparation:
(1) Put a final concentration of 1.32mM PA and 2.64mM OA into 10% BSA (free fatty acid free) DMEM cell culture medium.
(2) Shaking in the 37℃ incubator for one night.
Cell treatment:
(1) After cells form a uniform layer in the 96-well plate, aspirate the old FBA 10% cell culture medium;
(2) Replace with working solution 3mM free fatty acid, 10% BSA DMEM cell culture medium
(3) The control group is without 3mM free fatty acid, only the 10% BSA DMEM medium.
(4) Cell culture for 24 h
By this way, I have two question:
(1) the concentration of 10% BSA seems too high concentration?
(2) Could I change by 1% BSA and 10% FBS in DMEM cell culture medium. Some papers also used the FBS in their treatment. If so, how to evaluate the effect of fatty acid in FBS.
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