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Isolation of Blood-vessel-derived Multipotent Precursors from Human Skeletal Muscle
Published on: August 21, 2014
Isolation of blood-vessel-derived multipotent precursors from human skeletal muscle
William C W Chen1, Arman Saparov2, Mirko Corselli3
1Stem Cell Research Center, Department of Bioengineering and Orthopedic Surgery, University of Pittsburgh; chc88@pitt.edu.
Abstract:
Since the discovery of mesenchymal stem/stromal cells (MSCs), the native identity and localization of MSCs have been obscured by their retrospective isolation in culture. Recently, using fluorescence-activated cell sorting (FACS), we and other researchers prospectively identified and purified three subpopulations of multipotent precursor cells associated with the vasculature of human skeletal muscle. These three cell populations: myogenic endothelial cells (MECs), pericytes (PCs), and adventitial cells (ACs), are localized respectively to the three structural layers of blood vessels: intima, media, and adventitia. All of these human blood-vessel-derived stem cell (hBVSC) populations not only express classic MSC markers but also possess mesodermal developmental potentials similar to typical MSCs. Previously, MECs, PCs, and ACs have been isolated through distinct protocols and subsequently characterized in separate studies. The current isolation protocol, through modifications to the isolation process and adjustments in the selective cell surface markers, allows us to simultaneously purify all three hBVSC subpopulations by FACS from a single human muscle biopsy. This new method will not only streamline the isolation of multiple BVSC subpopulations but also facilitate future clinical applications of hBVSCs for distinct therapeutic purposes.
Insights
Researchers identified three distinct human blood-vessel-derived stem cell (hBVSC) populations in skeletal muscle using fluorescence-activated cell sorting. This new method enables simultaneous isolation of these multipotent cells for potential therapeutic applications.
Area of Science:
- Stem Cell Biology
- Vascular Biology
- Regenerative Medicine
Background:
- Mesenchymal stem/stromal cells (MSCs) native identity is unclear due to culture-based isolation.
- Previous research identified three distinct vascular cell populations (MECs, PCs, ACs) in human skeletal muscle.
- These populations were isolated and characterized separately using different protocols.
Purpose of the Study:
- To develop a streamlined method for simultaneously isolating three human blood-vessel-derived stem cell (hBVSC) subpopulations.
- To facilitate future clinical applications of hBVSCs.
Main Methods:
- Utilized fluorescence-activated cell sorting (FACS) for prospective cell identification and purification.
- Modified isolation protocols and adjusted cell surface markers for simultaneous purification.
- Isolated three hBVSC subpopulations (MECs, PCs, ACs) from human skeletal muscle biopsies.
Main Results:
- Successfully purified three distinct hBVSC subpopulations (MECs, PCs, ACs) from a single muscle biopsy.
- Demonstrated that these hBVSCs express classic MSC markers.
- Confirmed mesodermal developmental potentials similar to typical MSCs.
Conclusions:
- A novel FACS-based method allows simultaneous isolation of multiple hBVSC subpopulations.
- This streamlined approach simplifies the acquisition of therapeutically relevant stem cells.
- The findings pave the way for diverse clinical applications of hBVSCs.
