Chemically defined cytokine-free expansion of human haematopoietic stem cells
Chemically defined cytokine-free expansion of human haematopoietic stem cells
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DOI：
10.1038/s41586-023-05739-9
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发表时间：
2023-02-22
期刊：
影响因子：
64.8
通讯作者：
Yamazaki, Satoshi
中科院分区：
文献类型：
--
作者：
Sakurai, Masatoshi;Ishitsuka, Kantaro;Yamazaki, Satoshi
造血干细胞（HSC）是一种罕见的细胞类型，在移植后重建整个血液和免疫系统，可用作各种血液疾病的治愈性细胞疗法（1，2）。然而，体内HSC数量少使得生物学分析和临床应用都很困难，并且人HSC可以离体扩增的有限程度仍然是HSC移植更广泛和更安全的治疗用途的实质性障碍（3）。尽管已经测试了各种试剂以尝试刺激人HSC的扩增，但长期以来一直认为细胞因子对于离体支持HSC是必不可少的（4）。在这里，我们报告了一个培养系统的建立，允许长期离体扩增的人造血干细胞，通过完全取代外源性细胞因子和白蛋白与化学激动剂和基于己内酰胺的聚合物。磷酸肌醇3-激酶激活剂与血小板生成素受体激动剂和嘧啶并吲哚衍生物UM 171组合足以刺激能够在异种移植测定中连续植入的脐带血HSC的扩增。离体HSC扩增进一步通过分裂克隆移植测定和单细胞RNA测序分析得到支持。我们的化学成分确定的扩增培养系统将有助于推进临床HSC治疗。
Haematopoietic stem cells (HSCs) are a rare cell type that reconstitute the entire blood and immune systems after transplantation and can be used as a curative cell therapy for a variety of haematological diseases(1,2). However, the low number of HSCs in the body makes both biological analyses and clinical application difficult, and the limited extent to which human HSCs can be expanded ex vivo remains a substantial barrier to the wider and safer therapeutic use of HSC transplantation(3). Although various reagents have been tested in attempts to stimulate the expansion of human HSCs, cytokines have long been thought to be essential for supporting HSCs ex vivo(4). Here we report the establishment of a culture system that allows the long-term ex vivo expansion of human HSCs, achieved through the complete replacement of exogenous cytokines and albumin with chemical agonists and a caprolactam-based polymer. A phosphoinositide 3-kinase activator, in combination with a thrombopoietin-receptor agonist and the pyrimidoindole derivative UM171, were sufficient to stimulate the expansion of umbilical cord blood HSCs that are capable of serial engraftment in xenotransplantation assays. Ex vivo HSC expansion was further supported by split-clone transplantation assays and single-cell RNA-sequencing analysis. Our chemically defined expansion culture system will help to advance clinical HSC therapies.