title: CD146/MCAM defines functionality of human bone marrow stromal stem cell populations.
abstract: BACKGROUND
  Identification of surface markers for prospective isolation of functionally homogenous populations of human skeletal (stromal, mesenchymal) stem cells (hMSCs) is highly relevant for cell therapy protocols. Thus, we examined the possible use of CD146 to subtype a heterogeneous hMSC population.
  
  
  METHODS
  Using flow cytometry and cell sorting, we isolated two distinct hMSC-CD146(+) and hMSC-CD146(-) cell populations from the telomerized human bone marrow-derived stromal cell line (hMSC-TERT). Cells were examined for differences in their size, shape and texture by using high-content analysis and additionally for their ability to differentiate toward osteogenesis in vitro and form bone in vivo, and their migrational ability in vivo and in vitro was investigated.
  
  
  RESULTS
  In vitro, the two cell populations exhibited similar growth rate and differentiation capacity to osteoblasts and adipocytes on the basis of gene expression and protein production of lineage-specific markers. In vivo, hMSC-CD146(+) and hMSC-CD146(-) cells formed bone and bone marrow organ when implanted subcutaneously in immune-deficient mice. Bone was enriched in hMSC-CD146(-) cells (12.6 % versus 8.1 %) and bone marrow elements enriched in implants containing hMSC-CD146(+) cells (0.5 % versus 0.05 %). hMSC-CD146(+) cells exhibited greater chemotactic attraction in a transwell migration assay and, when injected intravenously into immune-deficient mice following closed femoral fracture, exhibited wider tissue distribution and significantly increased migration ability as demonstrated by bioluminescence imaging.
  
  
  CONCLUSIONS
  Our studies demonstrate that CD146 defines a subpopulation of hMSCs capable of bone formation and in vivo trans-endothelial migration and thus represents a population of hMSCs suitable for use in clinical protocols of bone tissue regeneration.
authors.affiliation: Department of Endocrinology and Metabolism, Molecular Endocrinology Laboratory (KMEB), Odense University Hospital, University of Southern Denmark, Winslowparken 25.1, 5000, Odense C, Denmark. lharkness@health.sdu.dk.
authors.forename: Linda
authors.initials: L
authors.lastname: Harkness
authors.affiliation: Department of Endocrinology and Metabolism, Molecular Endocrinology Laboratory (KMEB), Odense University Hospital, University of Southern Denmark, Winslowparken 25.1, 5000, Odense C, Denmark. wzaher@health.sdu.dk.|Stem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, 4852 Ash Shaikh Hasan Ibn Abdullah Al Ash Shaikh, Riyadh, 11461, Kingdom of Saudi Arabia. wzaher@health.sdu.dk.
authors.forename: Walid
authors.initials: W
authors.lastname: Zaher
authors.affiliation: Department of Endocrinology and Metabolism, Molecular Endocrinology Laboratory (KMEB), Odense University Hospital, University of Southern Denmark, Winslowparken 25.1, 5000, Odense C, Denmark. nditzel@health.sdu.dk.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Ditzel
authors.affiliation: Department of Endocrinology and Metabolism, Molecular Endocrinology Laboratory (KMEB), Odense University Hospital, University of Southern Denmark, Winslowparken 25.1, 5000, Odense C, Denmark. adiba.isa@gmail.com.
authors.forename: Adiba
authors.initials: A
authors.lastname: Isa
authors.affiliation: Department of Endocrinology and Metabolism, Molecular Endocrinology Laboratory (KMEB), Odense University Hospital, University of Southern Denmark, Winslowparken 25.1, 5000, Odense C, Denmark. mkassem@health.sdu.dk.|Danish Stem Cell Center (DanStem), Panum Institute, University of Copenhagen, Blegdamsvej 3B, Copenhagen, 2200, Denmark. mkassem@health.sdu.dk.|Stem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, 4852 Ash Shaikh Hasan Ibn Abdullah Al Ash Shaikh, Riyadh, 11461, Kingdom of Saudi Arabia. mkassem@health.sdu.dk.
authors.forename: Moustapha
authors.initials: M
authors.lastname: Kassem
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001861:Bone Regeneration; D051929:CD146 Antigen / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D048430:Cell Shape; D002470:Cell Survival; D002633:Chemotaxis; D029968:Escherichia coli Proteins; D005260:Female; D015870:Gene Expression; D006801:Humans; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D016688:Mice, Inbred NOD; D010006:Osteoblasts / Q000378:metabolism; D044968:Regenerative Medicine; D057705:Transendothelial and Transepithelial Migration
chemical_list: D015415:Biomarkers; D051929:CD146 Antigen; D029968:Escherichia coli Proteins; C497902:MCAM protein, human; C560213:lysis protein, E coli