Department of Microbiology and Immunology, The University of Melbourne and The Peter Doherty Institute for Infection and Immunity, Melbourne, VIC, Australia.
Singapore Immunology Network (SIgN), A*STAR (Agency for Science, Technology and Research), Singapore, Biopolis, Singapore.
Methods Mol Biol. 2021;2308:151-161. doi: 10.1007/978-1-0716-1425-9_13.
Hematopoiesis is a central process and is essential for the replenishment of short-lived leukocytes such as neutrophils. However, the molecular events underlining the developmental transition of quiescent hematopoietic stem cells into downstream progenitors and mature blood cells are not completely understood. Here, we describe the intrafemoral delivery of hematopoietic progenitors as a method to trace their development and differentiation lineage patterns within the bone marrow (BM) niche. Unlike other approaches, the direct adoptive transfer of progenitors into the BM cavity does not require prior irradiation preconditioning of recipient mice, and enables the delivery of lower cell numbers into the marrow space in a minimally perturbed environment. As a demonstrative example, we provide a protocol for the isolation of granulocyte-monocyte progenitors (GMP) by cell sorting, the delivery of these cells into recipient animals by intrafemoral transfer, and finally, the analysis of GMP-derived progenies by flow cytometry.
造血是一个核心过程，对于补充寿命较短的白细胞（如中性粒细胞）至关重要。然而，静止造血干细胞向下游祖细胞和成熟血细胞发育转变的分子事件尚不完全清楚。在这里，我们描述了通过股骨内输注造血祖细胞的方法，以追踪它们在骨髓（BM）龛内的发育和分化谱系模式。与其他方法不同，直接将祖细胞过继转移到 BM 腔中不需要预先对受体小鼠进行照射预处理，并且能够在最小干扰的环境中向骨髓空间中输送较少数量的细胞。作为一个示范示例，我们提供了一种通过细胞分选分离粒细胞-单核细胞祖细胞（GMP）的方案，通过股骨内转移将这些细胞递送到受体动物体内，最后通过流式细胞术分析 GMP 衍生的后代。
