Dissociation and inDrops microfluidic encapsulation of human gut tissues for single-cell atlasing studies

Autor: Alan J. Simmons, Ken S. Lau
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: STAR Protocols, Vol 3, Iss 3, Pp 101570- (2022)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2022.101570
Popis: Summary: In droplet-based single-cell RNA-sequencing (scRNA-seq) experiments, cells, along with some of their surrounding buffer and ambient material, are encapsulated into droplets for mRNA capture and barcoding. This protocol details the steps for human gut tissue dissociation using cold active protease, and subsequent isolation of single epithelial cells, with enrichment of viability through washes. Next, the steps for encapsulation on the inDrops scRNA-seq platform are described. This procedure has been demonstrated to be applicable to polyps, cancers, and inflamed tissues.For complete details on the use and execution of this protocol, please refer to Chen et al. (2021). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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