Hematopoietic stem cells retain functional potential and molecular identity in hibernation cultures.
Hematopoietic stem cells retain functional potential and molecular identity in hibernation cultures.
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DOI：
10.1016/j.stemcr.2021.04.002
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发表时间：
2021-06-08
影响因子：
5.9
通讯作者：
Kent DG
中科院分区：
文献类型：
--
作者：
Oedekoven CA;Belmonte M;Bode D;Hamey FK;Shepherd MS;Che JLC;Boyd G;McDonald C;Belluschi S;Diamanti E;Bastos HP;Bridge KS;Göttgens B;Laurenti E;Kent DG
单个造血干细胞（HSC）的分离和基因表达谱分析的进展已经允许其分子程序的深入解析。然而，长期HSC只能从成年小鼠骨髓中分离到接近纯度，从而排除了在不同生理状态下对其分子程序的研究。在这里，我们描述了一个强大的7天HSC冬眠培养系统，保持HSC作为单细胞在没有一个物理小生境。与新鲜分离的HSC相比，单个冬眠的HSC在集落形成能力和移植到初级和次级受体中方面保留了完整的功能潜力。冬眠HSC分子谱与其新鲜分离的对应物的比较显示出惊人的分子相似性，进一步解决了HSC自我更新的核心分子机制，同时还确定了可能影响HSC功能的关键因素，包括AP 1复合物（Jun，Fos和Ncor 2），Sult 1a 1和Cish的成员。最后，我们提供的证据表明，冬眠的小鼠HSC可以被转导，而不损害其自我更新活性，并证明了冬眠培养对人类HSC的适用性。在缺乏造血小生境的情况下维持完全功能性的单个HSC scRNA-seq鉴定HSC冬眠和SCF活化的分子调节剂人HSC活性可以保留在冬眠培养物中单个HSC可以在冬眠期间用慢病毒构建体转导在该研究中，大卫肯特和ElisaLaurenti的研究小组表明，完全功能的小鼠和人类造血干细胞（HSC）可以保存在非-在它们的原生微环境之外划分状态。单细胞移植和RNA测序证明了与新鲜分离的HSC的高度相似性，但也确定了参与HSC弹性的分子网络。
Advances in the isolation and gene expression profiling of single hematopoietic stem cells (HSCs) have permitted in-depth resolution of their molecular program. However, long-term HSCs can only be isolated to near purity from adult mouse bone marrow, thereby precluding studies of their molecular program in different physiological states. Here, we describe a powerful 7-day HSC hibernation culture system that maintains HSCs as single cells in the absence of a physical niche. Single hibernating HSCs retain full functional potential compared with freshly isolated HSCs with respect to colony-forming capacity and transplantation into primary and secondary recipients. Comparison of hibernating HSC molecular profiles to their freshly isolated counterparts showed a striking degree of molecular similarity, further resolving the core molecular machinery of HSC self-renewal while also identifying key factors that are potentially dispensable for HSC function, including members of the AP1 complex (Jun, Fos, and Ncor2), Sult1a1 and Cish. Finally, we provide evidence that hibernating mouse HSCs can be transduced without compromising their self-renewal activity and demonstrate the applicability of hibernation cultures to human HSCs. Fully functional single HSCs are maintained in the absence of a hematopoietic niche scRNA-seq identifies the molecular regulators of HSC hibernation and SCF activation Human HSC activity can be retained in hibernation cultures Single HSCs can be transduced with lentiviral constructs during hibernation In this study, the groups of David Kent and Elisa Laurenti show that fully functional mouse and human hematopoietic stem cells (HSCs) can be kept in a non-dividing state outside of their native microenvironment. Single-cell transplantations and RNA sequencing demonstrate a high degree of similarity to freshly isolated HSCs, but also identify molecular networks involved in HSC resilience.
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