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Updated: Jul 3, 2025

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从血液构成器官中隔离的流体细胞

Trine Kristiansen1, Christina Mayerhofer1, Karin Gustafsson2

  • 1Center for Regenerative Medicine, Massachusetts General Hospital; Harvard Stem Cell Institute; Department of Stem Cell and Regenerative Biology, Harvard University.

Journal of visualized experiments : JoVE
|February 12, 2024
PubMed
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这项研究详细介绍了从免疫器官中分离高质量的树皮细胞以进行单细胞测序的协议. 这些方法有助于分析健康,压力和衰老中的细胞异质性.

科学领域:

  • 免疫学 免疫学 免疫学
  • 细胞生物学 细胞生物学
  • 基因组学就是基因组学.

背景情况:

  • 单细胞测序揭示了血液构成器官的细胞异质性.
  • 了解干细胞种群是研究免疫系统功能,衰老和疾病的关键.
  • 以前的方法缺乏详细的协议来隔离特定的树皮细胞类型.

研究的目的:

  • 提供强大的,步骤明智的协议,以隔离来自小鼠和人类胸腺以及小鼠骨髓的高质量树皮细胞.
  • 为了优化树皮细胞的隔离,使其与单细胞多组学相兼容.
  • 引导研究人员在 stromal 细胞分析中取得可重现的结果.

主要方法:

  • 从小鼠和人类胸腺和骨髓中分离 stromal 细胞的详细方案的开发.
  • 优化样品消化和血液构造系的枯竭.
  • 光激活细胞分类 (FACS) 的应用用于细胞净化.
  • 评估孤立细胞是否适用于单细胞多组学.

主要成果:

  • 已建立的协议产生高质量的树皮细胞,适合单细胞测序.
  • 证明了消化和血统枯竭对树皮细胞产量和质量的影响.
  • 提供了FACS概况,说明了成功和不成功的分离技术.

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  • 在测序后的分析中展示了下游的树皮细胞产量.

    • 结论:

    • 优化的协议对于高质量的树皮细胞隔离至关重要.
    • 仔细考虑样本处理对于可重复的单细胞多组数据至关重要.
    • 这些协议将推进研究血液构造器官中树皮细胞异质性的研究.