[박상규] Foreskin fibroblast proliferation assay

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Foreskin fibroblast proliferation assay

 

For thymidine incorporation assay, Cells(FF: 3X10⁴⁾were seeded on 24 well plate and grown in DMEM medium supplemented with 10% fetal bovine serum and 50ug/ml streptomycin and penicillin in a 5% CO2 incubator at 37 degree for 12hrs. Culture medium was changed to serum free media with 1% streptomycin and penicillin without fetal bovine serum and incubated for 3hrs. p43 was treated as indicated concentration and cultured for 12hrs again. ³H labeled thymidine was treated with 1uci per well and incorporated to cells for 3 hrs. Cells were fixed with 5% TCA for 10 min and washed with 1X PBS 3 times. Cells were lysed by 0.5N NaOH and counted with Liquid scintillation counter.

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