无饲养层分化检测法用于分离培养小鼠造血干/祖细胞

Feeder-Free Differentiation Assay for Mouse Hematopoietic Stem and Progenitor Cells.

机构信息

Université de Paris, Institut de Recherche Saint-Louis, EMiLy, INSERM U1160, Paris, France.

Université de Paris, Institut de Recherche Saint-Louis, OPALE Carnot Institute, EMiLy, INSERM U1160, Paris, France.

出版信息

Methods Mol Biol. 2021;2308:47-58. doi: 10.1007/978-1-0716-1425-9_4.

DOI:10.1007/978-1-0716-1425-9_4

PMID:34057713

Abstract

Hematopoietic stem cells (HSCs) are responsible for replenishing immune cells and reside in bone marrow (BM) niches, which provide all cellular and molecular components required for their lifelong maintenance and differentiation. Although HSCs have been extensively analyzed and characterized, their ex vivo expansion, which constitutes a promising approach for therapeutic development in regenerative medicine, remains challenging. Here, we describe an original in vitro system allowing to quantify by flow cytometry the differentiation of mouse HSCs into lineage-primed multipotent hematopoietic progenitors (MPPs) in a cytokine-supplemented feeder-free medium.

摘要

造血干细胞（HSCs）负责补充免疫细胞，并存在于骨髓（BM）龛位中，这些龛位提供了其终生维持和分化所需的所有细胞和分子成分。尽管 HSCs 已经被广泛分析和表征，但它们的体外扩增仍然是再生医学治疗开发中的一个具有挑战性的问题。在这里，我们描述了一个原始的体外系统，该系统允许通过流式细胞术定量检测小鼠 HSCs 在细胞因子补充的无饲养层培养基中向谱系定型多能造血祖细胞（MPPs）的分化。