Cancer and Blood Disease Institute, Children's Hospital Los Angeles, Los Angeles, CA 90027, USA.
Department of Pediatrics, Keck School of Medicine, University of Southern California, Los Angeles, CA 90027, USA.
STAR Protoc. 2020 Aug 24;1(2):100090. doi: 10.1016/j.xpro.2020.100090. eCollection 2020 Sep 18.
Single cell RNA sequencing of human thymic cells is dependent on isolation of highly pure and viable cell populations. This protocol describes the isolation of CD34 progenitor and more differentiated CD34 fractions from post-natal thymic tissue to study thymopoiesis. CD34 cells represent <1% of thymic cells, so this protocol uses magnetic- followed by fluorescence-activated cell separation to isolate highly enriched CD34 cells. For complete details on the use and execution of this protocol, please refer to Le et al. (2020).
单细胞 RNA 测序依赖于高度纯和活细胞群体的分离。本方案描述了从出生后胸腺组织中分离 CD34 祖细胞和更分化的 CD34 亚群,以研究胸腺发生。CD34 细胞占胸腺细胞的<1%,因此本方案使用磁珠-随后进行荧光激活细胞分离来分离高度富集的 CD34 细胞。有关此方案的使用和执行的完整详细信息,请参考 Le 等人(2020 年)。