Isolation of Blood-vessel-derived Multipotent Precursors from Human Skeletal Muscle

William C.W. Chen; Arman Saparov; Mirko Corselli; Mihaela Crisan; Bo Zheng; Bruno P&#233;ault; Johnny Huard

doi:10.3791/51195

JoVE Journal
Biology

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JoVE Journal Biology

Isolation of Blood-vessel-derived Multipotent Precursors from Human Skeletal Muscle

从人骨骼肌隔离血管源性多能前体

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10:52 min

August 21, 2014

DOI: 10.3791/51195-v

William C.W. Chen¹, Arman Saparov^2,3, Mirko Corselli⁴, Mihaela Crisan⁵, Bo Zheng⁶, Bruno Péault^7,8, Johnny Huard⁹

¹Stem Cell Research Center, Department of Bioengineering and Orthopedic Surgery,University of Pittsburgh, ²Department of Orthopedic Surgery,University of Pittsburgh, ³Nazarbayev University Research and Innovation System,Nazarbayev University, ⁴Department of Orthopaedic Surgery, UCLA Orthopaedic Hospital and the Orthopaedic Hospital Research Center,University of California at Los Angeles, ⁵Department of Cell Biology,Erasmus MC Stem Cell Institute, ⁶OHSU Center for Regenerative Medicine,Oregon Health & Science University, ⁷Centre for Cardiovascular Science and MRC Centre for Regenerative Medicine,Queen's Medical Research Institute and University of Edinburgh, ⁸David Geffen School of Medicine and the Orthopaedic Hospital Research Center,University of California at Los Angeles, ⁹Stem Cell Research Center, Department of Orthopedic Surgery and McGowan Institute for Regenerative Medicine,University of Pittsburgh

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Overview

This article describes a method for isolating and purifying multipotent precursor cells from human blood vessels in skeletal muscle. The procedure allows for the simultaneous purification of myogenic endothelial cells, pericytes, and adventitial cells from different structural layers of blood vessels.

Key Study Components

Area of Science

Regenerative medicine
Cell biology
Stem cell research

Background

Human skeletal muscle contains multi-lineage precursor populations.
These precursor cells have potential applications in regenerative therapies.
Isolation of these cells can enhance understanding of their properties.
Blood vessels serve as a source for these multipotent cells.

Purpose of Study

To isolate specific subsets of blood vessel-derived precursor cells.
To purify myogenic endothelial cells, pericytes, and adventitial cells.
To analyze the purity and characteristics of these cell populations.

Methods Used

Dissociation of skeletal muscle biopsy using collagenase digestion.
Labeling of single cell suspension with fluorescence-conjugated antibodies.
Fluorescence-activated cell sorting (FACS) by flow cytometry.
Post-sort analysis including flow cytometry and RT-PCR.

Main Results

Successful isolation of three distinct multipotent precursor cell populations.
High purity of sorted cell subtypes confirmed through analysis.
Method demonstrates potential for studying the origin of mesenchymal stem cells.
Provides a foundation for future regenerative applications.

Conclusions

The described method effectively isolates precursor cells from human skeletal muscle.
These cells can be utilized for further research in regenerative medicine.
Understanding these populations may lead to advancements in therapeutic strategies.