The HEK293T, BV2, and NIH3T3 cells were purchased from iCell Bioscience (Shanghai, China). These cells were tested for mycoplasma contamination before experiments and they were all negative. These cell lines and primary microglia were all expanded in the following culture medium containing: Dulbecco’s modified Eagle’s medium (DMEM, Hyclone), 10% fetal bovine serum (FBS) (Gibco), 1x Pen/Strep (Gibco), and 1x MEM non-essential amino acids (Gibco). The cell incubator provided an environment of 5% CO2 and 37 °C.
Cerebral microglia were isolated from P0-P2 mice as previously described [44 (link)]. The forebrain was dissected out of the pia mater and then digested. The mixed cell suspension was incubated in the above-mentioned medium in 5% CO2 at 37 °C. After 14 days, the suspending cells (microglia) were rolled down and incubated for further experiments.
Cerebral microglia were isolated from P0-P2 mice as previously described [44 (link)]. The forebrain was dissected out of the pia mater and then digested. The mixed cell suspension was incubated in the above-mentioned medium in 5% CO2 at 37 °C. After 14 days, the suspending cells (microglia) were rolled down and incubated for further experiments.
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