Single cell sorting identifies progenitor cell population from full thickness bovine articular cartilage

Y Yu; H Zheng; J A Buckwalter; J A Martin

doi:10.1016/j.joca.2014.07.002

Single cell sorting identifies progenitor cell population from full thickness bovine articular cartilage

Osteoarthritis Cartilage. 2014 Sep;22(9):1318-26. doi: 10.1016/j.joca.2014.07.002. Epub 2014 Jul 16.

Authors

Y Yu¹, H Zheng², J A Buckwalter³, J A Martin⁴

Affiliations

¹ Department of Orthopaedics and Rehabilitation, The University of Iowa, Iowa City, IA, USA; Department of Biomedical Engineering, The University of Iowa, Iowa City, IA, USA. Electronic address: yin-yu@uiowa.edu.
² Department of Orthopaedics and Rehabilitation, The University of Iowa, Iowa City, IA, USA. Electronic address: Hongjun-zheng@uiowa.edu.
³ Department of Orthopaedics and Rehabilitation, The University of Iowa, Iowa City, IA, USA; Veterans Affairs Medical Center, Iowa City, IA, USA. Electronic address: joseph-buckwalter@uiowa.edu.
⁴ Department of Orthopaedics and Rehabilitation, The University of Iowa, Iowa City, IA, USA; Department of Biomedical Engineering, The University of Iowa, Iowa City, IA, USA. Electronic address: james-martin@uiowa.edu.

Abstract

Objective: To date, no approved clinical intervention successfully prevents the progressive degradation of injured articular cartilage that leads to osteoarthritis (OA). Stem/progenitor cell populations within tissues of diarthrodial joint have shown their therapeutic potential in treating OA. However, this potential has not been fully realized due in part to the heterogeneity of these subpopulations. Characterization of clonal populations derived from a single cell may help identify more homogenous stem/progenitor populations within articular cartilage. Moreover, chondrogenic potential of clonal populations from different zones could be further examined to elucidate their differential roles in maintaining articular cartilage homeostasis.

Method: We combined Fluorescence-activated cell sorting (FACS) and clonogenicity screening to identify stem/progenitor cells cloned from single cells. High-efficiency colony-forming cells (HCCs) were isolated, and evaluated for stem/progenitor cell characteristics. HCCs were also isolated from different zones of articular cartilage. Their function was compared by lineage-specific gene expression, and differentiation potential.

Results: A difference in colony-forming efficiency was observed in terms of colony sizes. HCCs were highly clonogenic and multipotent, and overexpressed stem/progenitor cell markers. Also, proliferation and migration associated genes were over-expressed in HCCs. HCCs showed zonal differences with deep HCCs more chondrogenic and osteogenic than superficial HCCs.

Conclusion: Our approach is a simple yet practical way to identify homogeneous stem/progenitor cell populations with clonal origin. The discovery of progenitor cells demonstrates the intrinsic self-repairing potential of articular cartilage. Differences in differentiation potential may represent the distinct roles of superficial and deep zone stem/progenitor cells in the maintenance of articular cartilage homeostasis.

Keywords: Articular cartilage; Single cell; Stem/progenitor cell.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Cartilage, Articular / cytology*
Cartilage, Articular / metabolism
Cattle
Cell Differentiation / physiology
Cell Proliferation / genetics
Cell Separation / methods
Cells, Cultured
Chemotaxis / genetics
Chemotaxis / physiology
Chondrocytes / cytology
Chondrocytes / metabolism
Chondrogenesis / genetics
Chondrogenesis / physiology
Colony-Forming Units Assay
Flow Cytometry / methods
Gene Expression
Multipotent Stem Cells / cytology
Stem Cells / cytology*
Stem Cells / metabolism

Grants and funding

P50 AR055533/AR/NIAMS NIH HHS/United States