Step 4: DO NOT SHAKE
Notes: Simple routine, minimal irritation risk
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E., Siegel, P
Cells were seeded on collagen-coated plates (250 g/ml rat collagen, Roche) at a density of 150,000 cells/cm 2 and cultivated for 3 h in Williams E medium (PAN Biotech) containing penicillin (100 U/ml, Gibco), streptomycin (0.1 mg/ml, Gibco), gentamycin (10 M, PAN Biotech), dexamethasone (100 nM, Sigma), stable l-glutamine (2 mM, Sigma), insulin supplement (2 ng/ml, Sigma), and 10% Sera Plus (PAN Biotech)