P-EMBL-77 - P-EMBL-77
The murine fetal liver cell line AFT024 (a kind gift from I. R. Lemischka, Princeton University, Princeton) was maintained in DMEM (Gibco, Grand Island, N.Y., USA) supplemented with 20% FCS, 50 µM 2-Mercaptoethanol (Bio-Rad, Hercules, CA) as described before. Cells were grown to confluency in 24- or 96-well plates precoated with 0.1% gelatin (Specialty Media, Lavette, NJ) and maintained at 33C. AFT024 cells were then irradiated with 2000 rads using a irradiator 24 hours prior to using the feeder layer. Media was then changed to RPMI supplemented with 20% FCS (R20), Penicillin 1000 U/ml, Streptomycin 100 U/ml, and 50 µmol/l 2ME. Medium was supplemented with Flf3-Ligand (FL) 10 ng/ml and Thrombopoietin (Tpo) 10 ng/ml.
max temperature, media, min temperature, start Time, stop Time