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- 积分
- 460
- 威望
- 460
- 包包
- 1467
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背景文献如下: C+ @2 d% f0 [; c! p% |7 r5 ?
For osteogenic differentiation, fourth passage MSC were
o) z" N5 p! T5 W, P7 ?plated at 5×103 cells/cm2 in a 9.6-cm2 slideflask (Nunc," k5 s o8 s/ V n5 k
Denmark), grown up to 80% confluence, and incubated in5 l1 w+ h* i* ?0 [+ H
osteogenic medium (NH Osteodiff Medium, Miltenyi4 n$ x4 }: p: W V# c& a
Biotec, Germany). The medium was replaced every 3–4 days. After 10 days, cultures were washed with PBS and/ ^. m# Y& v9 B5 i* r+ k
fixed in a solution of ice-cold 70% ethanol for 10 min. For$ N1 K* \- z0 C% Q" r3 s
alkaline phosphatase staining, slides were fixed in 100%
- f$ {3 g6 j* v8 b6 A; |methanol for 5 min, washed with PBS, and then incubated4 ?3 ?; j% y0 ?
with 5-bromo-4-chloro3-indolyl phosphate/nitro blue tetrazolium
9 z9 s: e# I6 a- E" C, @(BCIP-NBT, Sigma-Aldrich) for 15–30 min, strictly
( b5 h* R/ k0 a2 Z v$ f8 r, Dfollowing manufacturer’s instructions |
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发表于 2010-3-15 20:30
发表于 2010-3-31 11:24
