从非孕和孕鼠宫颈组织中分离上皮细胞用于单细胞 RNA 测序的方案。
Protocol to dissociate epithelia from non-pregnant and pregnant mouse cervical tissue for single-cell RNA-sequencing.
机构信息
Department of Obstetrics and Gynecology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; Cecil H. and Ida Green Center for Reproductive Biology Sciences, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
Cecil H. and Ida Green Center for Reproductive Biology Sciences, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
出版信息
STAR Protoc. 2023 Dec 15;4(4):102631. doi: 10.1016/j.xpro.2023.102631. Epub 2023 Oct 31.
A challenge in studying cervical epithelial cell biology at the single-cell level is that differentiated subtypes, in particular mucus-secreting goblet cells, are sensitive to disassociating enzymes making isolation of all epithelial subpopulations difficult. Here we present a protocol to dissociate epithelia from non-pregnant and pregnant mouse cervical tissue for single-cell RNA-sequencing. We describe steps for harvesting cervices, preparing cervical tissue, dissociation of cervical cells, and viability checks. We then detail library preparation, sequencing, and procedure for data analysis. For complete details on the use and execution of this protocol, please refer to Cooley et al. (2023)..
研究宫颈上皮细胞生物学的一个挑战是，在单细胞水平上，分化亚型，特别是分泌粘液的杯状细胞，对分离酶很敏感，这使得所有上皮亚群的分离都很困难。在这里，我们介绍了一种从非怀孕和怀孕小鼠宫颈组织中分离上皮细胞进行单细胞 RNA 测序的方案。我们描述了收获宫颈、准备宫颈组织、分离宫颈细胞和活力检查的步骤。然后，我们详细介绍了文库制备、测序以及数据分析程序。有关此方案的使用和执行的完整详细信息，请参阅 Cooley 等人。(2023)。
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