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Split sub-confluent cultures (70-80%) 1:4 to 1:10 seeding at approximately 3 x 10⁴ cells/cm² using 0.25% trypsin or trypsin/EDTA; 5% CO₂; 37 °C. It is important to inactivate the trypsin with an equal volume of trypsin inhibitor (the culture medium contains too low a serum content to inactivate the trypsin). To remove all traces of trypsin and trypsin inhibitor pellet the cells by centrifugation and resuspend the cells in fresh medium. Media change every second day.
Stemline Keratinocyte Medium II (Sigma S0196) + Stemline Keratinocyte Growth Supplement (Sigma S9945) + 2 mM Glutamine + 2% Fetal Bovine Serum (FBS). Addition of 2% serum was found to increase cell viability.
Prostate cancer moderately well differentiated, early stage
STR-PCR Data: Amelogenin: X,Y CSF1PO: 12 D13S317: 11 D16S539: 11,14 D5S818: 12,13 D7S820: 7,10 THO1: 7,9 TPOX: 8 vWA: 17,19
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