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本帖最后由 细胞海洋 于 2015-3-18 10:03 编辑
1 }7 c+ e6 h& z$ n8 A! R7 n0 S. Z1 ]% s( D: N* r
Carboxyfl uorescein succinimidyl ester (CFSE;0 S8 @' g# h" T1 p1 |/ D
Invitrogen)-labeled PBMC were cultured at3 v; j/ j( q+ w; x3 N$ |2 P
400 000 cells/well in a 24-well plate with cultured,- `# P% H! w# N
freshly thawed or paraformaldehyde (PFA)-fi xed1 U0 Y9 D% s- | s1 u, V2 }
MSC at a ratio of MSC:PBMC of 1:3 and 1:9. T
6 j0 a2 v( w; m, p3 v3 \# \/ Rlymphocytes were stimulated using 0.4 μ g/mL antihuman+ n6 B7 j& ]2 k# G6 Q( i
CD3 and CD28 antibodies (clone HIT3a6 V: w$ i9 p- o: Z4 W, U
and CD28.2, respectively, from BD Pharmagen,+ n8 a: z' P! H. t
Mississauga, ON, Canada). T-cell proliferation was
# g$ E) ^' j5 s4 N0 B* Fdetermined 4 days later by fl ow cytometry analysis
0 V1 D* I' |' ?% v; N! U% s& ]of CFSE fl uorescence intensity.
[- h. A# u1 n
" o& R/ |4 k, e# n* J5 _/ Z% j6 dparaformaldehyde (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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