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Can Arthroscopically Harvested Synovial Stem Cells Be Preferentially Sorted Using Stage-Specific Embryonic Antigen 4 Antibody for Cartilage, Bone, and Adipose Regeneration?

  • Jingting Li
    Affiliations
    Stem Cell and Tissue Engineering Laboratory, Department of Orthopaedics, West Virginia University, Morgantown, West Virginia, U.S.A.

    Department of Exercise Physiology, West Virginia University, Morgantown, West Virginia, U.S.A.
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  • Douglas D. Campbell
    Affiliations
    Stem Cell and Tissue Engineering Laboratory, Department of Orthopaedics, West Virginia University, Morgantown, West Virginia, U.S.A.
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  • George K. Bal
    Affiliations
    Stem Cell and Tissue Engineering Laboratory, Department of Orthopaedics, West Virginia University, Morgantown, West Virginia, U.S.A.
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  • Ming Pei
    Correspondence
    Address correspondence to Ming Pei, M.D., Ph.D., Stem Cell and Tissue Engineering Laboratory, Department of Orthopaedics, West Virginia University, PO Box 9196, One Medical Center Drive, Morgantown, WV 26506-9196, U.S.A.
    Affiliations
    Stem Cell and Tissue Engineering Laboratory, Department of Orthopaedics, West Virginia University, Morgantown, West Virginia, U.S.A.

    Department of Exercise Physiology, West Virginia University, Morgantown, West Virginia, U.S.A.

    Department of Mechanical & Aerospace Engineering, West Virginia University, Morgantown, West Virginia, U.S.A.
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      Purpose

      The aim of this study was to investigate the relation between stage-specific embryonic antigen 4 (SSEA4) expression and synovium-derived stem cell (SDSC) lineage differentiation.

      Methods

      Human SDSCs were collected during arthroscopic surgery from 4 young patients with anterior cruciate ligament injuries. Passage 2 SDSCs were sorted by fluorescence-activated cell sorting using phycoerythrin-conjugated monoclonal antibody against SSEA4 into 3 groups: SSEA4(+) cells, SSEA4(−) cells, and unsorted control cells. After 1 more passage, expanded cells from each group were evaluated for SSEA4 expression by use of flow cytometry as well as multilineage differentiation capacities, including chondrogenesis, adipogenesis, and osteogenesis, using biochemical analysis, histologic analysis, immunostaining, and real-time polymerase chain reaction.

      Results

      After cell sorting, 1 more passage expansion decreased SSEA4(+) cells from 99.8% to 79.2% and increased SSEA4(−) cells from 4.4% to 53.3% compared with 70.3% in the unsorted cell population. SSEA4(−) SDSCs with a lower cell proliferation exhibited higher chondrogenic potential (in terms of the ratio of glycosaminoglycan to DNA [P < .001] and COL2A1 [type II collagen] messenger RNA [mRNA] [P < .001]) and adipogenic potential (in terms of oil red O staining and quantitative assay [P = .007], LPL [lipoprotein lipase] mRNA [P = .005], and CEBP [CCAAT/enhancer-binding protein alpha] mRNA [P = .010]). In contrast, SSEA4(+) SDSCs retained cell expansion and enhanced osteogenic capacity, as evidenced by intense calcium deposition stained by alizarin red S and a significantly elevated expression of OPN (osteopontin) mRNA (P = .007).

      Conclusions

      In this study, for the first time, we showed the benefit of using the surface marker SSEA4 in SDSCs to preferentially sort a mixed population of cells. SSEA4(+) SDSCs indicated a strong potential for osteogenesis rather than chondrogenesis and adipogenesis.

      Clinical Relevance

      SDSC-based mesenchymal tissue regeneration can be easily achieved by arthroscopic harvesting followed by quick cell sorting.
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