Long-term ex vivo expansion of mouse hematopoietic stem cells
Long-term ex vivo expansion of mouse hematopoietic stem cells
复制标题
DOI：
10.1038/s41596-019-0263-2
复制
发表时间：
2020-01-08
期刊：
影响因子：
14.8
通讯作者：
Yamazaki, Satoshi
中科院分区：
文献类型：
--
作者：
Wilkinson, Adam C.;Ishida, Reiko;Yamazaki, Satoshi
利用多能和自我更新能力，造血干细胞(HSCs)可以在一生中维持造血。然而，这种非凡能力背后的机制仍未被发现，至少部分原因是由于HSCs的缺乏，以及HSCs在含有定义不明确的白蛋白补充剂(如牛血清白蛋白)的培养液中适度的体外扩张。在这里，我们描述了一个简单的平台，用于在完全定义的无白蛋白条件下体外扩增1个月的功能性小鼠造血干细胞。该培养系统在一个月的时间内可提供236至899倍的扩增，并可对HSC异质性进行克隆分析。大量扩增的造血干细胞可以使造血干细胞移植到非条件受者体内，否则这在常规上是不可行的，因为需要大量的造血干细胞。因此，该方案提供了一种强大的方法，用于询问HSC的自我更新和谱系承诺，更广泛地，研究和表征造血和免疫系统。使用完全定义的无白蛋白培养系统，功能性小鼠造血干细胞(HSCs)体外扩增236至899倍。此外，还介绍了HSC异质性的克隆分析和HSC移植。
Utilizing multipotent and self-renewing capabilities, hematopoietic stem cells (HSCs) can maintain hematopoiesis throughout life. However, the mechanism behind such remarkable abilities remains undiscovered, at least in part because of the paucity of HSCs and the modest ex vivo expansion of HSCs in media that contain poorly defined albumin supplements such as bovine serum albumin. Here, we describe a simple platform for the expansion of functional mouse HSCs ex vivo for >1 month under fully defined albumin-free conditions. The culture system affords 236- to 899-fold expansion over the course of a month and is also amenable to clonal analysis of HSC heterogeneity. The large numbers of expanded HSCs enable HSC transplantation into nonconditioned recipients, which is otherwise not routinely feasible because of the large numbers of HSCs required. This protocol therefore provides a powerful approach with which to interrogate HSC self-renewal and lineage commitment and, more broadly, to study and characterize the hematopoietic and immune systems.Functional mouse hematopoietic stem cells (HSCs) are expanded 236- to 899-fold ex vivo using a fully defined albumin-free culture system. Clonal analysis of HSC heterogeneity and HSC transplantation are also described.