8×105 cells at different passages were pelleted into a micromass by centrifugation at 450g for 10 minutes in a 15-ml conical polypropylene tube and cultured in complete medium or chondrogenic medium at 37° C, 5% CO2, which contained low-glucose Dulbecco modified Eagle medium (Gibco, Invitrogen corporation, Carlsbad, US), supplemented with 10 ng/ml transforming growth factor (TGF)-β3 (R&D, Minneapolis, MN, US), 500 ng/ml bone morphogenetic protein (BMP)-2 (R&D Systems, Inc., Minneapolis, MN, US), 10-7M dexamethasone, 50 μg/ml ascorbate-2-phosphate, 40 μg/ml proline, 100 μg/ml pyruvate (all from Sigma-Aldrich, St Louis, MO), and 1:100 diluted ITS+Premix (6.25 mg/ml insulin, 6.25 mg/ml transferrin, 6.25 mg/ml selenous acid, 1.25 mg/ml bovine serum albumin, 5.35 mg/ml linoleic acid) (Becton Dickinson, Franklin Lakes, US). , @! ~' U1 J* ?
I used this mothod in my human BMSC and the results were good. 7 |: p3 |- W: O0 c8 g2 i: y6 NKeep the reagent fresh and store -20° C.& X- s4 O9 N) P& h( b
You can refer my paper[attach]41019[/attach]作者: 柠檬草 时间: 2012-4-18 00:04