Protocol for the induction of hindlimb ischemia and isolation of muscle endothelial cells in mice

Min Seong Kim; Shantel Vital; Changwon Park

doi:10.1016/j.xpro.2022.102017

Protocol for the induction of hindlimb ischemia and isolation of muscle endothelial cells in mice

STAR Protoc. 2023 Mar 17;4(1):102017. doi: 10.1016/j.xpro.2022.102017. Epub 2023 Jan 12.

Authors

Min Seong Kim¹, Shantel Vital², Changwon Park³

Affiliations

¹ Department of Molecular and Cellular Physiology, School of Graduate Studies, Louisiana State University Health, Shreveport, LA 71103, USA.
² CCDS Surgical Models Core, Louisiana State University Health, Shreveport, LA 71103, USA.
³ Department of Molecular and Cellular Physiology, School of Graduate Studies, Louisiana State University Health, Shreveport, LA 71103, USA. Electronic address: changwon.park@lsuhs.edu.

Abstract

A mouse model of hindlimb ischemia is an important tool for studying diverse therapeutic approaches for vascularization with high surgical success and low mortality rates. Here, we present a protocol for the induction of hindlimb ischemia in mice, including the surgery procedure and steps to analyze blood perfusion in the ischemic area using a laser speckle contrast analyzer. We also detail the isolation of endothelial cells from thigh muscles using flow cytometry after ischemic surgery. For complete details on the use and execution of this protocol, please refer to Park et al. (2016).¹.

Keywords: Cell Biology; Cell Isolation; Flow Cytometry/Mass Cytometry; Health Sciences; Model Organisms.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Endothelial Cells*
Hindlimb
Ischemia / therapy
Mice
Muscle, Skeletal
Neovascularization, Physiologic* / physiology

Grants and funding

R01 HL119291/HL/NHLBI NIH HHS/United States