title: Multipotential human adipose-derived stromal stem cells exhibit a perivascular phenotype in vitro and in vivo.
abstract: Mesenchymal stem-like cells identified in different tissues reside in a perivascular niche. In the present study, we investigated the putative niche of adipose-derived stromal/stem cells (ASCs) using markers, associated with mesenchymal and perivascular cells, including STRO-1, CD146, and 3G5. Immunofluorescence staining of human adipose tissue sections, revealed that STRO-1 and 3G5 co-localized with CD146 to the perivascular regions of blood vessels. FACS was used to determine the capacity of the CD146, 3G5, and STRO-1 specific monoclonal antibodies to isolate clonogenic ASCs from disassociated human adipose tissue. Clonogenic fibroblastic colonies (CFU-F) were found to be enriched in those cell fractions selected with either STRO-1, CD146, or 3G5. Flow cytometric analysis revealed that cultured ASCs exhibited similar phenotypic profiles in relation to their expression of cell surface markers associated with stromal cells (CD44, CD90, CD105, CD106, CD146, CD166, STRO-1, alkaline phosphatase), endothelial cells (CD31, CD105, CD106, CD146, CD166), haematopoietic cells (CD14, CD31, CD45), and perivascular cells (3G5, STRO-1, CD146). The immunoselected ASCs populations maintained their characteristic multipotential properties as shown by their capacity to form Alizarin Red positive mineralized deposits, Oil Red O positive lipid droplets, and Alcian Blue positive proteoglycan-rich matrix in vitro. Furthermore, ASCs cultures established from either STRO-1, 3G5, or CD146 selected cell populations, were all capable of forming ectopic bone when transplanted subcutaneously into NOD/SCID mice. The findings presented here, describe a multipotential stem cell population within adult human adipose tissue, which appear to be intimately associated with perivascular cells surrounding the blood vessels.
authors.affiliation: Myeloma Research Program, Bone and Cancer Laboratories, Division of Haematology, Institute of Medical and Veterinary Science, Hanson Institute and University of Adelaide, Adelaide, Australia.
authors.forename: A C W
authors.initials: AC
authors.lastname: Zannettino
authors.forename: S
authors.initials: S
authors.lastname: Paton
authors.forename: A
authors.initials: A
authors.lastname: Arthur
authors.forename: F
authors.initials: F
authors.lastname: Khor
authors.forename: S
authors.initials: S
authors.lastname: Itescu
authors.forename: J M
authors.initials: JM
authors.lastname: Gimble
authors.forename: S
authors.initials: S
authors.lastname: Gronthos
mesh_terms: D050156:Adipogenesis; D000273:Adipose Tissue / Q000166:cytology*; D000328:Adult; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D051929:CD146 Antigen / Q000378:metabolism; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D020219:Chondrogenesis; D018076:DNA, Complementary / Q000096:biosynthesis; D005260:Female; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D019084:Fluorescent Antibody Technique, Indirect; D005456:Fluorescent Dyes; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D016513:Mice, SCID; D010012:Osteogenesis; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017154:Stromal Cells / Q000166:cytology* / Q000637:transplantation; D014183:Transplantation, Heterologous
chemical_list: D000911:Antibodies, Monoclonal; D015415:Biomarkers; D051929:CD146 Antigen; D018076:DNA, Complementary; D005456:Fluorescent Dyes; D016650:Fluorescein-5-isothiocyanate