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Generation and immortalization of MEFs

JC Jiali Cai
DH Die Hu
JS Judy Sakya
TS Tao Sun
DW Daoyong Wang
LW Lin Wang
XM Xiaohua Mao
ZS Zhenyi Su
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Abin-1+/– mice were bred with Abin-1+/–Ripk1D138N or Abin-1+/–Ripk3–/– mice, and pregnancy was terminated at the E11-13 stage. Embryos were homogenized individually and treated with trypsin/EDTA, sieved through a 70-µm filter and primary MEFs were cultured in high-glucose DMEM supplemented with 15% FBS, nonessential amino acids, sodium pyruvate, penicillin, streptomycin, and amphotericin B. At passages 4–6, primary MEFs were immortalized by transfection with SV40 small + large T antigen-expressing plasmid (Addgene, Cat# 22298) and keeping culturing for a few generations.

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