分选出Tfh细胞后体外培养

shiyisyl

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。