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- 积分
- 460
- 威望
- 460
- 包包
- 1467
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背景文献如下:( y& |8 Y+ I3 V% N) E5 X
For osteogenic differentiation, fourth passage MSC were
# m0 ^3 z- D) A9 L5 V7 Iplated at 5×103 cells/cm2 in a 9.6-cm2 slideflask (Nunc,2 \! z1 f o( t1 a. v6 k2 x- k4 R5 C
Denmark), grown up to 80% confluence, and incubated in
8 r6 \& J- Y4 B2 N" z# V7 \osteogenic medium (NH Osteodiff Medium, Miltenyi2 o+ W" Z& c3 Y9 l+ t% ~1 X
Biotec, Germany). The medium was replaced every 3–4 days. After 10 days, cultures were washed with PBS and6 Q! ^0 n2 L F7 D4 h! |
fixed in a solution of ice-cold 70% ethanol for 10 min. For
: ]! r+ e4 A) T4 C4 N Halkaline phosphatase staining, slides were fixed in 100%0 I+ A$ X6 R) p
methanol for 5 min, washed with PBS, and then incubated3 z+ J8 E* Q& d5 y. o! ]0 O' t
with 5-bromo-4-chloro3-indolyl phosphate/nitro blue tetrazolium
* W x: m3 s }* d9 f2 X7 Y(BCIP-NBT, Sigma-Aldrich) for 15–30 min, strictly
7 e$ @! ~1 N% A }/ ufollowing manufacturer’s instructions |
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发表于 2010-3-15 20:30
发表于 2010-3-31 11:24
