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本帖最后由 细胞海洋 于 2015-3-18 10:03 编辑 + t1 \/ a) O V. X- k7 g
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Carboxyfl uorescein succinimidyl ester (CFSE;5 \# b& f& y, L& q' `/ {: \
Invitrogen)-labeled PBMC were cultured at3 C! a" {- J: z& m) m
400 000 cells/well in a 24-well plate with cultured,& v, G6 p; M6 G8 k% c1 `- s
freshly thawed or paraformaldehyde (PFA)-fi xed' H6 N9 Q! p; S' G
MSC at a ratio of MSC:PBMC of 1:3 and 1:9. T9 g) W/ p2 d6 w; q$ |
lymphocytes were stimulated using 0.4 μ g/mL antihuman
2 a4 R" J5 p- W2 E3 E( d; t+ ^& xCD3 and CD28 antibodies (clone HIT3a2 u7 J# X8 }: e) N( {
and CD28.2, respectively, from BD Pharmagen,
! |# p t. S9 \/ e5 ]Mississauga, ON, Canada). T-cell proliferation was* ` ] W$ o$ L( }( x9 I
determined 4 days later by fl ow cytometry analysis
7 _4 ]: P" h) c# x, Cof CFSE fl uorescence intensity.
( I; v8 k/ P- m t& X0 r0 H$ K& d: H) I) E4 u- z
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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