Department of Medicine/Hematology Oncology, Indiana University School of Medicine, Indianapolis, IN, USA.
Methods Mol Biol. 2023;2567:181-189. doi: 10.1007/978-1-0716-2679-5_12.
Mesenchymal stromal cells (MSCs) are the crucial component of the hematopoietic stem and progenitor cell (HSPC) niche in the bone marrow. Therefore, an ex vivo culture system that recapitulates the marrow microenvironment is important to understanding the niche's regulatory role on HSPC function and improving ex vivo HSPC expansion for clinical transplantation. Herein, a procedure for ex vivo expansion of MSCs from human bone marrow cells and their identification and characterization is described. In addition, a protocol for MSC and HSPC coculture assay is presented. This MSC-HSPC coculture assay can be used for ex vivo expansion of HSPC. Furthermore, this assay is also useful for qualitative analysis of MSCs capable of supporting hematopoiesis.
间充质基质细胞(MSCs)是骨髓中造血干细胞和祖细胞(HSPC)龛的关键组成部分。因此,体外重现骨髓微环境的培养系统对于理解龛在 HSPC 功能上的调节作用以及改善临床移植用 HSPC 的体外扩增非常重要。本文描述了从人骨髓细胞中体外扩增 MSCs 的程序及其鉴定和特征。此外,还介绍了 MSC 和 HSPC 共培养测定法的方案。这种 MSC-HSPC 共培养测定法可用于 HSPC 的体外扩增。此外,该测定法还可用于定性分析支持造血的 MSC。