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CD90 (Thy-1)-positive selection enhances osteogenic capacity of human adipose-derived stromal cells.

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  • Additional Information
    • Source:
      Publisher: Mary Ann Liebert, Inc Country of Publication: United States NLM ID: 101466659 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1937-335X (Electronic) Linking ISSN: 19373341 NLM ISO Abbreviation: Tissue Eng Part A Subsets: MEDLINE
    • Publication Information:
      Original Publication: New Rochelle, NY : Mary Ann Liebert, Inc.
    • Subject Terms:
      Osteogenesis*; Adipose Tissue/*cytology ; Thy-1 Antigens/*metabolism; Adult ; Alkaline Phosphatase/metabolism ; Animals ; Cell Differentiation ; Cell Size ; Female ; Flow Cytometry ; Gene Expression Regulation ; Humans ; Mice ; Middle Aged ; Skull/pathology ; Staining and Labeling ; Stromal Cells/cytology ; Stromal Cells/enzymology
    • Abstract:
      Background: Stem cell-based bone tissue engineering with adipose-derived stromal cells (ASCs) has shown great promise for revolutionizing treatment of large bone deficits. However, there is still a lack of consensus on cell surface markers identifying osteoprogenitors. Fluorescence-activated cell sorting has identified a subpopulation of CD105(low) cells with enhanced osteogenic differentiation. The purpose of the present study was to compare the ability of CD90 (Thy-1) to identify osteoprogenitors relative to CD(105).
      Methods: Unsorted cells, CD90(+), CD90(-), CD105(high), and CD105(low) cells were treated with an osteogenic differentiation medium. For evaluation of in vitro osteogenesis, alkaline phosphatase (ALP) staining and alizarin red staining were performed at 7 days and 14 days, respectively. RNA was harvested after 7 and 14 days of differentiation, and osteogenic gene expression was examined by quantitative real-time polymerase chain reaction. For evaluation of in vivo osteogenesis, critical-sized (4-mm) calvarial defects in nude mice were treated with the hydroxyapatite-poly(lactic-co-glycolic acid) scaffold seeded with the above-mentioned subpopulations. Healing was followed using micro-CT scans for 8 weeks. Calvaria were harvested at 8 weeks postoperatively, and sections were stained with Movat's Pentachrome.
      Results: Transcriptional analysis revealed that the CD90(+) subpopulation was enriched for a more osteogenic subtype relative to the CD105(low) subpopulation. Staining at day 7 for ALP was greatest in the CD90(+) cells, followed by the CD105(low) cells. Staining at day 14 for alizarin red demonstrated the greatest amount of mineralized extracellular matrix in the CD90(+) cells, again followed by the CD105(low) cells. Quantification of in vivo healing at 2, 4, 6, and 8weeks postoperatively demonstrated increased bone formation in defects treated with CD90(+) ASCs relative to all other groups. On Movat's Pentachrome-stained sections, defects treated with CD90(+) cells showed the most robust bony regeneration. Defects treated with CD90(-) cells, CD105(high) cells, and CD105(low) cells demonstrated some bone formation, but to a lesser degree when compared with the CD90(+) group.
      Conclusions: While CD105(low) cells have previously been shown to possess an enhanced osteogenic potential, we found that CD90(+) cells are more capable of forming bone both in vitro and in vivo. These data therefore suggest that CD90 may be a more effective marker than CD105 to isolate a highly osteogenic subpopulation for bone tissue engineering.
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    • Grant Information:
      R01 DE021683 United States DE NIDCR NIH HHS; United States HHMI Howard Hughes Medical Institute; R01-DE019434 United States DE NIDCR NIH HHS; R01-DE021683-01 United States DE NIDCR NIH HHS
    • Accession Number:
      0 (Thy-1 Antigens)
      EC 3.1.3.1 (Alkaline Phosphatase)
    • Publication Date:
      Date Created: 20121211 Date Completed: 20130926 Latest Revision: 20211021
    • Publication Date:
      20231215
    • Accession Number:
      PMC3589870
    • Accession Number:
      10.1089/ten.TEA.2012.0370
    • Accession Number:
      23216074