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Guidelines for the Isolation and Characterization of Murine Vascular Smooth Muscle Cells. A Report from the International Society of Cardiovascular Translational Research

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Abstract

Vascular smooth muscle cells (VSMCs) play important roles in cardiovascular disorders and biology. Outlined in this paper is a step-by-step procedure for isolating aortic VSMCs from adult C57BL6J male mice by enzymatic digestion of the aorta using collagenase. The plating, culturing, and subculturing of the isolated cells are discussed in detail along with techniques to characterize VSMC phenotype by gene expression and immunofluorescence. Traction force microscopy was used to characterize contractility of single subcultured VSMCs at baseline.

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Abbreviations

Acta2:

Smooth muscle α-actin

APS:

Ammonium persulfate

Cald1:

Caldesmon1

Cnn1:

Calponin1

EdU:

5-Ethynyl-2′-deoxyuridine

FN:

Fibronectin

Myh11:

Smooth muscle myosin heavy chain

P1:

Passage 1

P2:

Passage 2

PA:

Polyacrylamide

PDMS:

Polydimethylsiloxane

sm22α (Tagln):

Smooth muscle22α (also known as Transgelin (Tagln))

Smtn:

Smoothelin

TFM:

Traction force microscopy

TEMED:

Tetramethylethylenediamine

VSMC:

Vascular smooth muscle cell

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Acknowledgments

The authors wish to thank the University of Minnesota Genomics Core for single-cell RNA sequencing.

Source of Funding

The sources of funding are from DK54733 (LW), DK60521 (LW), LHI Innovation Grant (LW, JLH), and the Lindahl Foundation (JLH).

The Use of Laboratory Animals

All institutional and national guidelines for the care and use of laboratory animals were followed and approved by the appropriate institutional committees.

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Authors and Affiliations

  1. Division of Cardiology, Lillehei Heart Institute, Department of Medicine, University of Minnesota, 4-138 CCRB, 2231 6th Street SE, Minneapolis, MN, 55455, USA

    Neeta Adhikari, Kadambari Chandra Shekar, Rodney Staggs & Jennifer L. Hall

  2. Department of Bioengineering, University of Minnesota, Minneapolis, MN, USA

    Zaw Win, Kerianne Steucke & Patrick Alford

  3. Department of Pharmacology, University of Minnesota, Minneapolis, MN, USA

    Yi-Wei Lin & Li-Na Wei

Authors
  1. Neeta Adhikari
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  2. Kadambari Chandra Shekar
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  3. Rodney Staggs
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  4. Zaw Win
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  5. Kerianne Steucke
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  6. Yi-Wei Lin
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  7. Li-Na Wei
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  8. Patrick Alford
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  9. Jennifer L. Hall
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Corresponding author

Correspondence to Jennifer L. Hall.

Additional information

Associate Editor Lorrie Kirshenbaum oversaw the review of this article

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Adhikari, N., Shekar, K.C., Staggs, R. et al. Guidelines for the Isolation and Characterization of Murine Vascular Smooth Muscle Cells. A Report from the International Society of Cardiovascular Translational Research. J. of Cardiovasc. Trans. Res. 8, 158–163 (2015). https://doi.org/10.1007/s12265-015-9616-6

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  • DOI: https://doi.org/10.1007/s12265-015-9616-6

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