各位老师,请教个问题。
想要将小鼠A 生发中心的Tfh细胞分选出来,再和小鼠B 的GC B细胞共同体外培养,目的是:检测小鼠A的Tfh对小鼠B的GC B细胞的作用效果。但是Tfh细胞的体外培养并维持Tfh细胞,应该加什么细胞因子呢?是否想需要DC细胞?
看过了倪老师的另外一个帖子,但是帖子中是对CD4+T诱导Tfh细胞。而我想做的是:体外培养并维持Tfh。请教各位如何操作。
目前已经查阅的文献如下:
B cells (4×10^4/well) were plated alone or with2 × 10^4 act.caspase1⁺ Tfh cells or 2 × 10^4 act. caspase-1⁻ Tfh cells in the presence of 2 μg/ml soluble anti-CD3(Biolegend) and 5 μg/ml anti-IgM (JacksonImmunoresearch Laboratories) for 6 days. Atday 6, cells were collected and stained with fluorophore-conjugated Ab for flowcytometry. Activated B cells (CD19+GL-7+) were detected by flow cytometry. (ZhaoZ, Xu B, Wang S, et.al. Tfh cells with NLRP3inflammasome activation are essential for high-affinity antibody generation,germinal centre formation and autoimmunity[J]. Annals of the RheumaticDiseases, 2022, 81(7): 1006–1012.)
为正确培养Tfh细胞,同时防止Tfh向Th1转化(Tfh表型较多,想要维持GC-Tfh表型),该如何培养,请各位老师指教。
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