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

STAR Protocols (Dec 2023)

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

Affiliations

Qianglin Liu: School of Animal Sciences, AgCenter, Louisiana State University, Baton Rouge, LA, USA
Chaoyang Li: School of Animal Sciences, AgCenter, Louisiana State University, Baton Rouge, LA, USA
Yuxia Li: School of Animal Sciences, AgCenter, Louisiana State University, Baton Rouge, LA, USA
Leshan Wang: School of Animal Sciences, AgCenter, Louisiana State University, Baton Rouge, LA, USA
Xujia Zhang: School of Animal Sciences, AgCenter, Louisiana State University, Baton Rouge, LA, USA
Buhao Deng: Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China
Peidong Gao: School of Animal Sciences, AgCenter, Louisiana State University, Baton Rouge, LA, USA
Mohammad Shiri: Department of Computer Science, Old Dominion University, Norfolk, VA, USA
Fozi Alkaifi: Department of Computer Science, Old Dominion University, Norfolk, VA, USA
Junxing Zhao: Department of Animal Sciences, Shanxi Agricultural University, Taigu, Shanxi, China
Jacqueline M. Stephens: Pennington Biomedical Research Center, Baton Rouge, LA, USA; Department of Biological Sciences, Louisiana State University, Baton Rouge, LA, USA
Constantine A. Simintiras: School of Animal Sciences, AgCenter, Louisiana State University, Baton Rouge, LA, USA
Joseph Francis: Department of Comparative Biomedical Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA, USA
Jiangwen Sun: Department of Computer Science, Old Dominion University, Norfolk, VA, USA
Xing Fu: 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; Corresponding author

Journal volume & issue: Vol. 4, no. 4
p. 102703

Abstract

Read online

Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

About the journal

Abstract

Keywords