Single-cell isolation from full-thickness human intestinal tissue resections for single-cell RNA sequencing

被引：1

作者：

West, Gail A. ^{[1
]}

Zhao, Shuai ^{[1
]}

Nguyen, Quang Tam ^{[1
,2
]}

Christensen, Stephen M. ^{[3
]}

Gordon, Ilyssa O. ^{[2
,4
]}

Holubar, Stefan D. ^{[2
,5
]}

Kravarik, Kellie M. ^{[3
]}

Fiocchi, Claudio ^{[1
,6
]}

Mukherjee, Pranab K. ^{[1
,2
]}

Rieder, Florian ^{[1
,2
,6
]}

机构：

[1] Cleveland Clin, Lerner Res Inst, Dept Inflammat & Immun, Cleveland, OH 44195 USA

[2] Cleveland Clin, Ctr Global Translat Inflammatory Bowel Dis Res, Cleveland, OH 44195 USA

[3] Pfizer Inc, Worldwide Res Dev & Med, Cambridge, MA USA

[4] Cleveland Clin, Robert J Tomsich Pathol & Lab Med Inst, Dept Pathol, Cleveland, OH USA

[5] Cleveland Clin, Digest Dis & Surg Inst, Dept Colorectal Surg, Cleveland, OH USA

[6] Cleveland Clin, Digest Dis & Surg Inst, Dept Gastroenterol Hepatol & Nutr, Cleveland, OH 44195 USA

来源：

STAR PROTOCOLS | 2023年 / 4卷 / 04期

关键词：

Cell isolation; Cell separation/fractionation; RNA-seq; Single Cell;

D O I：

10.1016/j.xpro.2023.102686

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Single-cell isolation techniques allow the investigation of physical and functional relationships between individual cells within a complex cell population. Here, we present a protocol for single-cell isolation from full-thickness intestinal tissue resections. We describe steps for pre-processing specimens, isolation of lamina propria and muscular layers, and red blood cell lysis. We then detail fixation of isolated cells and assessment of cell quality. The resulting cell suspension can be subjected to RNA sequencing on the 103 Chromium platform.For complete details on the use and execution of this protocol, please refer to Mukherjee et al.1

引用

页数：14

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