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定量分析人类初始 B 细胞在体外细胞培养中的增殖动力学和分化。

Quantifying Human Naïve B Cell Proliferation Kinetics and Differentiation in Controlled In Vitro Cell Culture.

机构信息

Immunology Division, Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia.

Department of Medical Biology, University of Melbourne, Parkville, VIC, Australia.

出版信息

Methods Mol Biol. 2024;2826:167-187. doi: 10.1007/978-1-0716-3950-4_13.

Abstract

Division tracking dyes like Cell Trace Violet (CTV) enable the quantification of cell proliferation, division, and survival kinetics of human naïve B cell responses in vitro. Human naïve B cells exhibit distinct responses to different stimuli, with CpG and anti-Ig inducing a T cell-independent (TI) response, while CD40L and IL-21 promote a T cell-dependent (TD) response that induces isotype switching and differentiation into antibody-secreting cells (ASCs). Both stimulation methods yield valuable insights into the intrinsic programming of B cell health within individuals, making them useful for clinical investigations. For instance, quantitative analysis from these B cell populations could reveal biologically meaningful measurements such as the average number of division rounds and the time to cells' fate. Here, we describe a novel in vitro culture setup for CTV-labelled human naïve B cells and a method for obtaining precise time-based data on proliferation, division-linked isotype switching, and differentiation.

摘要

分裂追踪染料,如 CellTrace Violet(CTV),可用于量化体外人类初始 B 细胞反应的细胞增殖、分裂和存活动力学。人类初始 B 细胞对不同刺激表现出不同的反应,CpG 和抗 Ig 诱导非依赖 T 细胞(TI)反应,而 CD40L 和 IL-21 促进依赖 T 细胞(TD)反应,诱导同种型转换和分化为抗体分泌细胞(ASC)。这两种刺激方法都深入了解了个体内 B 细胞健康的内在编程,使其成为临床研究的有用工具。例如,这些 B 细胞群体的定量分析可以揭示具有生物学意义的测量结果,例如平均分裂轮数和细胞命运的时间。在这里,我们描述了一种新的体外 CTV 标记人类初始 B 细胞培养方案,以及一种获取增殖、与分裂相关的同种型转换和分化的精确基于时间的数据的方法。

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