Hi, I have been working with 3T3-L1 cells for over a year now.However, over the last couple of months they stopped differentiating into adipocytes.
I seed at 200k cells/well (12 well plate), and grow in 10% FBS, 1% P/S HG DMEM. Two days post confluence I induce with 0.5 mM IBMX, 1uM Dexa, and 1ug/ml insulin for two days. change to insulin media from 2 till day 4, and continue with 10% FBS, 1% P/S HG DMEM till day 8-10, refreshed every 2-3 days.
I have considered working with new IBMX, dexa, as well as new batch of cells. nothing seems to work.
can you please advice? thank you :)
I have attached some images 10 days after adipogenesis
Maybe traits and genes(if both recessive) can be reached through strong enough somatic mutations: https://www.researchgate.net/publication/380427514_Kalergi_and_Hart-Cellerand_Memetics_White_Antifragility
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