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本帖最后由 细胞海洋 于 2015-3-18 10:03 编辑 0 q3 I1 o+ a q, }% D9 H# r
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Carboxyfl uorescein succinimidyl ester (CFSE;& [4 }4 B0 _+ R9 J E" {
Invitrogen)-labeled PBMC were cultured at) r3 u& K! E1 O( _* L
400 000 cells/well in a 24-well plate with cultured,0 `& n) P5 {9 ?6 E3 Q
freshly thawed or paraformaldehyde (PFA)-fi xed* _. M4 m7 P( h
MSC at a ratio of MSC:PBMC of 1:3 and 1:9. T: e$ K) i# U! b, O6 q" Y
lymphocytes were stimulated using 0.4 μ g/mL antihuman
9 W* _+ v& Q O* e+ LCD3 and CD28 antibodies (clone HIT3a) H" ^. ~3 q) t
and CD28.2, respectively, from BD Pharmagen,0 f% C* w2 d$ C/ c/ ?2 d7 B
Mississauga, ON, Canada). T-cell proliferation was( J1 d1 a* {8 Z- B7 l
determined 4 days later by fl ow cytometry analysis
7 E N$ i5 c# g3 g- Q1 U) Wof CFSE fl uorescence intensity.
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paraformaldehyde (PFA)-fixed MSC 这是什么意思?记得共培养是用丝裂霉素处理? |
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