Isolation of Type I and Type II pericytes from mouse skeletal muscles

Abhijit Nirwane, Jyoti Gautam, Yao Yao

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

Pericytes are perivascular multipotent cells that show heterogeneity in different organs or even within the same tissue. In skeletal muscles, there are at least two pericyte subpopulations (called type I and type II), which express different molecular markers and have distinct differentiation capabilities. Using NG2-DsRed and Nestin-GFP double-transgenic mice, type I (NG2-DsRed+Nestin-GFP-) and type II (NG2-DsRed+Nestin-GFP+) pericytes have been successfully isolated. However, the availability of these double-transgenic mice prevents the widespread use of this purification method. This work describes an alternative protocol that allows for the easy and simultaneous isolation of type I and type II pericytes from skeletal muscles. This protocol utilizes the fluorescence-activated cell sorting (FACS) technique and targets PDGFRβ, rather than NG2, together with the Nestin-GFP signal. Following isolation, type I and type II pericytes show distinct morphologies. In addition, type I and type II pericytes isolated with this new method, like those isolated from the double-transgenic mice, are adipogenic and myogenic, respectively. These results suggest that this protocol can be used to isolate pericyte subpopulations from skeletal muscles and possibly from other tissues.

Original languageEnglish (US)
Article numbere55904
JournalJournal of Visualized Experiments
Volume2017
Issue number123
DOIs
StatePublished - May 26 2017

Bibliographical note

Funding Information:
This work was partially supported by a Fund-A-Fellow grant from the Myotonic Dystrophy Foundation (MDF-FF-2014-0013) and the Scientist Development Grant from the American Heart Association (16SDG29320001).

Publisher Copyright:
© 2017 Journal of Visualized Experiments.

Keywords

  • Adipogenesis
  • Developmental biology
  • FACS
  • Issue 123
  • Myogenesis
  • Nestin-GFP
  • PDGFRβ
  • Type I pericyte
  • Type II pericyte

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