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Clonal Analysis of Embryonic Hematopoietic Stem Cell Precursors Using Single Cell Index Sorting Combined with Endothelial Cell Niche Co-culture
Published on: May 8, 2018
Single cell sorting identifies progenitor cell population from full thickness bovine articular cartilage
Y Yu1, H Zheng2, J A Buckwalter3
1Department of Orthopaedics and Rehabilitation, The University of Iowa, Iowa City, IA, USA; Department of Biomedical Engineering, The University of Iowa, Iowa City, IA, USA.
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.
Insights
Researchers identified homogeneous stem/progenitor cells from articular cartilage using a novel method. These cells show potential for self-repair, with distinct zonal differences impacting cartilage homeostasis and osteoarthritis treatment.
Area of Science:
- Biomedical Engineering
- Regenerative Medicine
- Cartilage Biology
Background:
- Osteoarthritis (OA) is characterized by progressive articular cartilage degradation, with no current clinical interventions preventing its progression.
- Stem/progenitor cells in joints hold therapeutic promise for OA, but their heterogeneity hinders clinical application.
- Clonal analysis of single cells can identify homogenous stem/progenitor populations within articular cartilage.
Purpose of the Study:
- To develop a method for identifying homogenous stem/progenitor cell populations from articular cartilage.
- To characterize the stem/progenitor cell properties and chondrogenic potential of clonal populations from different articular cartilage zones.
- To elucidate the differential roles of superficial and deep zone progenitor cells in maintaining articular cartilage homeostasis.
Main Methods:
- Combined Fluorescence-activated cell sorting (FACS) and clonogenicity screening to isolate single cells.
- Identified and isolated high-efficiency colony-forming cells (HCCs) with stem/progenitor characteristics.
- Compared zonal differences in HCCs through lineage-specific gene expression and differentiation potential assays.
Main Results:
- A novel approach successfully identified homogenous, clonally derived stem/progenitor cell populations (HCCs).
- HCCs exhibited high clonogenicity, multipotency, and overexpressed stem/progenitor markers, along with proliferation and migration genes.
- Deep zone HCCs demonstrated greater chondrogenic and osteogenic potential compared to superficial zone HCCs, indicating zonal functional differences.
Conclusions:
- The developed method provides a practical means to isolate homogenous stem/progenitor cells of clonal origin.
- The identification of progenitor cells highlights the intrinsic self-repair capacity of articular cartilage.
- Zonal variations in differentiation potential suggest distinct functional roles for superficial and deep zone stem/progenitor cells in cartilage maintenance.
