Progenitor cell isolation from mouse epididymal adipose tissue and sequencing library construction

Qianglin Liu; Chaoyang Li; Yuxia Li; Leshan Wang; Xujia Zhang; Buhao Deng; Peidong Gao; Mohammad Shiri; Fozi Alkaifi; Junxing Zhao; Jacqueline M Stephens; Constantine A Simintiras; Joseph Francis; Jiangwen Sun; Xing Fu

doi:10.1016/j.xpro.2023.102703

Progenitor cell isolation from mouse epididymal adipose tissue and sequencing library construction

STAR Protoc. 2023 Dec 15;4(4):102703. doi: 10.1016/j.xpro.2023.102703. Epub 2023 Nov 9.

Authors

Affiliations

¹ School of Animal Sciences, AgCenter, Louisiana State University, Baton Rouge, LA, USA.
² Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.
³ Department of Computer Science, Old Dominion University, Norfolk, VA, USA.
⁴ Department of Animal Sciences, Shanxi Agricultural University, Taigu, Shanxi, China.
⁵ Pennington Biomedical Research Center, Baton Rouge, LA, USA; Department of Biological Sciences, Louisiana State University, Baton Rouge, LA, USA.
⁶ Department of Comparative Biomedical Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA, USA.
⁷ School of Animal Sciences, AgCenter, Louisiana State University, Baton Rouge, LA, USA; Department of Biological and Agricultural Engineering, Louisiana State University, Baton Rouge, LA, USA. Electronic address: xfu1@agcenter.lsu.edu.

Abstract

Here, we present a protocol to isolate progenitor cells from mouse epididymal visceral adipose tissue and construct bulk RNA and assay for transposase-accessible chromatin with sequencing (ATAC-seq) libraries. We describe steps for adipose tissue collection, cell isolation, and cell staining and sorting. We then detail procedures for both ATAC-seq and RNA sequencing library construction. This protocol can also be applied to other tissues and cell types directly or with minor modifications. For complete details on the use and execution of this protocol, please refer to Liu et al. (2023).¹.

Keywords: Cell-based Assays; Flow Cytometry; RNA-seq; Sequencing.

MeSH terms

Adipose Tissue*
Animals
Biological Assay*
Cell Movement
Mice
Sequence Analysis, RNA
Stem Cells

Abstract

MeSH terms

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