您好,请问:1.步骤5,为何要换新板?新板需要预先包被anti-CD3吗?2.步骤6,说是只加中和抗体不加细胞因子?我没...

lz
lin zhang
Feb 17, 2012
protocol Protocol: In vitro Differentiation of Mouse Th0, Th1 and Th2 from Naïve CD4 T Cells
您好,请问:
1.步骤5,为何要换新板?新板需要预先包被anti-CD3吗?
2.步骤6,说是只加中和抗体不加细胞因子?我没过这样的
3.这样刺激一轮的话阳性细胞的比例有多少?

谢谢您了!
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Fanglian He Answered Feb 25, 2012

Bio-protocol

The step 5 has been updated.
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Jia Li Author Answered Feb 24, 2012

SRI International

您好,
1,步骤5,因为anti-CD3的抗体会造成细胞死亡,所以转移到新板这一步是必须的,这也是为什么一开始anti-CD3需要plate-bound的原因,方便转板。如果用soluble anti-CD3的话就需要洗细胞。
2,步骤5,需要细胞因子。不好意思,已请编辑修改。
3,对WT control来说,Th0的话看不到IL-4和IL-17,有10-15%的IFN-gamma,Th1的大部分细胞都表达IFN-gamma,Th2和Th17有15-20%左右的阳性率。
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