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- 积分
- 460
- 威望
- 460
- 包包
- 1467
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背景文献如下:
2 t/ j$ U1 W+ e" j- @( gFor osteogenic differentiation, fourth passage MSC were
" ?3 C1 b6 X) @! qplated at 5×103 cells/cm2 in a 9.6-cm2 slideflask (Nunc,
# ~( |+ ~" h4 o2 H9 t, z. LDenmark), grown up to 80% confluence, and incubated in
4 Y( H: i; j- l4 L/ aosteogenic medium (NH Osteodiff Medium, Miltenyi
: P" }; S" F: B NBiotec, Germany). The medium was replaced every 3–4 days. After 10 days, cultures were washed with PBS and. M4 B+ V$ j* ^6 p
fixed in a solution of ice-cold 70% ethanol for 10 min. For
/ W9 I# r9 l1 W4 G: salkaline phosphatase staining, slides were fixed in 100%
% s2 q" [. c: M* M7 x5 Qmethanol for 5 min, washed with PBS, and then incubated
/ n6 i0 S! s/ ~% y4 {( D% Q3 Jwith 5-bromo-4-chloro3-indolyl phosphate/nitro blue tetrazolium7 Y6 _( h) l& w* }! C" t& }+ x* G
(BCIP-NBT, Sigma-Aldrich) for 15–30 min, strictly
( V' m9 X5 M1 pfollowing manufacturer’s instructions |
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发表于 2010-3-15 20:30
发表于 2010-3-31 11:24
