Perturbation of single hematopoietic stem cell fates in artificial niches.
Perturbation of single hematopoietic stem cell fates in artificial niches.
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DOI：
10.1039/b815718a
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发表时间：
2009-01
期刊：
影响因子：
--
通讯作者：
Blau HM
中科院分区：
文献类型：
--
作者：
Lutolf MP;Doyonnas R;Havenstrite K;Koleckar K;Blau HM
造血干细胞（hsc）在体内具有广泛的自我更新能力，并已成功地用于临床治疗造血恶性肿瘤，但由于在培养中这种自我更新能力丧失，供应有限。利用干细胞生物学和生物工程相结合的方法，我们描述了一种新的水凝胶微孔阵列平台，用于评估体内微环境或生态位特征的分泌或系留蛋白对体外HSC命运的影响。单细胞延时显微分析对于克服facs富集造血干细胞不可避免的异质性至关重要。针对特定蛋白（Wnt3a和N-Cadherin），微孔中单个造血干细胞的增殖动力学降低或异步分裂增加与小鼠体内随后的连续长期血液重构密切相关。在给定蛋白存在下分裂一次的单细胞能够在体内重建，这为造血干细胞在体外的自我更新分裂提供了证据。这些结果验证了水凝胶微孔平台是一种广泛适用的范例，可用于分析复杂干细胞生态位中特定信号的调节作用。
Hematopoietic stem cells (HSCs) are capable of extensive self-renewal in vivo and are successfully employed clinically to treat hematopoietic malignancies, yet are in limited supply as in culture this self-renewal capacity is lost. Using an approach at the interface of stem cell biology and bioengineering, here we describe a novel platform of hydrogel microwell arrays for assessing the effects of either secreted or tethered proteins characteristic of the in vivo microenvironment, or niche, on HSC fate in vitro. Time-lapse microscopic analyses of single cells were crucial to overcoming inevitable heterogeneity of FACS-enriched HSCs. A reduction in proliferation kinetics or an increase in asynchronous division of single HSCs in microwells in response to specific proteins (Wnt3a and N-Cadherin) correlated well with subsequent serial long-term blood reconstitution in mice in vivo. Single cells that divided once in the presence of a given protein were capable of in vivo reconstitution, providing evidence of self-renewal divisions of HSCs in vitro. These results validate the hydrogel microwell platform as a broadly applicable paradigm for dissecting the regulatory role of specific signals within a complex stem cell niche.
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