Michael Smith Laboratories, University of British Columbia, Vancouver, British Columbia, Canada.
Nat Methods. 2011 May 22;8(7):581-6. doi: 10.1038/nmeth.1614.
Heterogeneity in cell populations poses a major obstacle to understanding complex biological processes. Here we present a microfluidic platform containing thousands of nanoliter-scale chambers suitable for live-cell imaging studies of clonal cultures of nonadherent cells with precise control of the conditions, capabilities for in situ immunostaining and recovery of viable cells. We show that this platform mimics conventional cultures in reproducing the responses of various types of primitive mouse hematopoietic cells with retention of their functional properties, as demonstrated by subsequent in vitro and in vivo (transplantation) assays of recovered cells. The automated medium exchange of this system made it possible to define when Steel factor stimulation is first required by adult hematopoietic stem cells in vitro as the point of exit from quiescence. This technology will offer many new avenues to interrogate otherwise inaccessible mechanisms governing mammalian cell growth and fate decisions.
细胞群体的异质性是理解复杂生物过程的主要障碍。在这里，我们提出了一种包含数千个纳升级规模腔室的微流控平台，非常适合对非贴壁细胞的克隆培养进行活细胞成像研究，可精确控制条件、原位免疫染色能力和有活力细胞的回收。我们表明，该平台通过后续的体外和体内（移植）检测回收细胞的功能特性，模拟传统培养方法复制各种类型原始鼠造血细胞的反应。该系统的自动培养基交换使得可以确定 Steel 因子刺激何时首次需要体外成体造血干细胞退出静止状态。这项技术将为研究否则无法访问的哺乳动物细胞生长和命运决定的调控机制提供许多新途径。
