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本帖最后由 细胞海洋 于 2015-3-18 10:03 编辑
1 U8 K2 O2 s1 |( x
2 ^! M, [, M: VCarboxyfl uorescein succinimidyl ester (CFSE;5 T& B; ^5 q' M _5 T( R
Invitrogen)-labeled PBMC were cultured at& `. O+ f2 g4 k- T' g/ M8 m
400 000 cells/well in a 24-well plate with cultured,
7 o; e, ]7 P& i2 k! Ufreshly thawed or paraformaldehyde (PFA)-fi xed
2 o3 X6 p: Y% T: h& jMSC at a ratio of MSC:PBMC of 1:3 and 1:9. T
7 U G7 y1 j5 g$ olymphocytes were stimulated using 0.4 μ g/mL antihuman- }- D: j# y, w4 w( j
CD3 and CD28 antibodies (clone HIT3a* x5 L f' t+ F- E6 v
and CD28.2, respectively, from BD Pharmagen,$ M. h) B+ K7 p, ~' v
Mississauga, ON, Canada). T-cell proliferation was
6 n3 H3 e+ K1 Z" u, h! Mdetermined 4 days later by fl ow cytometry analysis w7 q: H# D) d4 ~8 ^% q# w. Y
of CFSE fl uorescence intensity.% {- v, Y! b: w* {1 Y
; r+ v `4 P+ Tparaformaldehyde (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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