The cell line 3A sub E was derived from 3A(tPA-30-1)after the original line started to senesce and went through crisis. 3A subE has an unlimited lifespan and expresses human chorionic gonadotropin (hCG)
Aspirate medium and add 0.25% trypsin/EDTA; incubate at room temperature until cells detach. Dilute with fresh medium and aliquot into new flasks at a split ratio 1:2. Renew fluid 1-2 times per week. Incubate culture at 37°C, 5% CO2 for initial 24 hour
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