CD146 expression on mesenchymal stem cells is associated with their vascular smooth muscle commitment

Bone marrow mesenchymal stem cells (MSCs) are plastic adherent cells that can differentiate into various tissue lineages, including osteoblasts, adipocytes and chondrocytes. However, this progenitor property is not shared by all cells within the MSC population. In addition, MSCs vary in their proliferation capacity and expression of markers. Because of heterogeneity of CD146 expression in the MSC population, we compared CD146−/Low and CD146High cells under clonal conditions and after sorting of the non‐clonal cell population to determine whether this expression is associated with specific functions. CD146−/Low and CD146High bone marrow MSCs did not differ in colony‐forming unit‐fibroblast number, osteogenic, adipogenic and chondrogenic differentiation or in vitro haematopoietic‐supportive activity. However, CD146−/Low clones proliferated slightly but significantly faster than did CD146High clones. In addition, a strong expression of CD146 molecule was associated with a commitment to a vascular smooth muscle cell (VSMC) lineage characterized by a strong up‐regulation of calponin‐1 and SM22α expression and an ability to contract collagen matrix. Thus, within a bone marrow MSC population, certain subpopulations characterized by high expression of CD146, are committed towards a VSMC lineage.

[1]  R. Class,et al.  Propagation and senescence of human marrow stromal cells in culture: a simple colony‐forming assay identifies samples with the greatest potential to propagate and differentiate , 1999, British journal of haematology.

[2]  K. Sobue,et al.  Expressional regulation of smooth muscle cell-specific genes in association with phenotypic modulation , 2004, Molecular and Cellular Biochemistry.

[3]  M. Ehinger,et al.  CD146 expression on primary nonhematopoietic bone marrow stem cells is correlated with in situ localization. , 2011, Blood.

[4]  K. Hirschi,et al.  PDGF, TGF-β, and Heterotypic Cell–Cell Interactions Mediate Endothelial Cell–induced Recruitment of 10T1/2 Cells and Their Differentiation to a Smooth Muscle Fate , 1998, The Journal of cell biology.

[5]  D. Prockop,et al.  An Alizarin red-based assay of mineralization by adherent cells in culture: comparison with cetylpyridinium chloride extraction. , 2004, Analytical biochemistry.

[6]  T. Ochi,et al.  Smooth Muscle Cell Phenotype-dependent Transcriptional Regulation of the α1 Integrin Gene* , 1997, The Journal of Biological Chemistry.

[7]  G. Owens,et al.  A transforming growth factor beta (TGFbeta) control element drives TGFbeta-induced stimulation of smooth muscle alpha-actin gene expression in concert with two CArG elements. , 1997, The Journal of biological chemistry.

[8]  J. Sampol,et al.  Identification of the S-Endo 1 endothelial-associated antigen. , 1996, Biochemical and biophysical research communications.

[9]  J. Loscalzo,et al.  Vascular Calcification: Pathobiological Mechanisms and Clinical Implications , 2006, Circulation research.

[10]  S. Gronthos,et al.  MUC18, a member of the immunoglobulin superfamily, is expressed on bone marrow fibroblasts and a subset of hematological malignancies , 1998, Leukemia.

[11]  A. Friedenstein,et al.  STROMAL CELLS RESPONSIBLE FOR TRANSFERRING THE MICROENVIRONMENT OF THE HEMOPOIETIC TISSUES: Cloning In Vitro and Retransplantation In Vivo , 1974, Transplantation.

[12]  Sebastian Brenner,et al.  Expression of the melanoma cell adhesion molecule in human mesenchymal stromal cells regulates proliferation, differentiation, and maintenance of hematopoietic stem and progenitor cells , 2013, Haematologica.

[13]  D. Prockop,et al.  Adult Bone Marrow Stem/Progenitor Cells (MSCs) Are Preconditioned by Microenvironmental "Niches" in Culture: A Two-Stage Hypothesis for Regulation of MSC Fate , 2005, Science's STKE.

[14]  J. Ringe,et al.  Specific plasma membrane protein phenotype of culture-amplified and native human bone marrow mesenchymal stem cells. , 2008, Blood.

[15]  M. Corselli,et al.  The tunica adventitia of human arteries and veins as a source of mesenchymal stem cells. , 2012, Stem cells and development.

[16]  Rodolfo Quarto,et al.  Ex vivo enrichment of mesenchymal cell progenitors by fibroblast growth factor 2. , 2003, Experimental cell research.

[17]  N. Kulagina,et al.  Fibroblast precursors in normal and irradiated mouse hematopoietic organs. , 1976, Experimental hematology.

[18]  M. Lichtman.  The relationship of stromal cells to hemopoietic cells in marrow. , 1984, Kroc Foundation series.

[19]  Joelle Dulong,et al.  Clinical-grade mesenchymal stromal cells produced under various good manufacturing practice processes differ in their immunomodulatory properties: standardization of immune quality controls. , 2013, Stem cells and development.

[20]  O. Bagasra,et al.  Cultured adherent cells from marrow can serve as long-lasting precursor cells for bone, cartilage, and lung in irradiated mice. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Y. Kato,et al.  Retention of multilineage differentiation potential of mesenchymal cells during proliferation in response to FGF. , 2001, Biochemical and biophysical research communications.

[22]  G. Owens,et al.  A Transforming Growth Factor β (TGFβ) Control Element Drives TGFβ-induced Stimulation of Smooth Muscle α-Actin Gene Expression in Concert with Two CArG Elements* , 1997, The Journal of Biological Chemistry.

[23]  Katie C. Russell,et al.  In Vitro High‐Capacity Assay to Quantify the Clonal Heterogeneity in Trilineage Potential of Mesenchymal Stem Cells Reveals a Complex Hierarchy of Lineage Commitment , 2010, Stem cells.

[24]  D. Schaffer,et al.  Transforming Growth Factor‐β and Notch Signaling Mediate Stem Cell Differentiation into Smooth Muscle Cells , 2010, Stem cells.

[25]  G. Crooks,et al.  Perivascular support of human hematopoietic stem/progenitor cells. , 2013, Blood.

[26]  G. Gabbiani,et al.  α-Smooth muscle actin is expressed in a subset of bone marrow stromal cells in normal and pathological conditions , 1989 .

[27]  M. Humbert,et al.  IgG from patients with pulmonary arterial hypertension and/or systemic sclerosis binds to vascular smooth muscle cells and induces cell contraction , 2011, Annals of the rheumatic diseases.

[28]  V. Koteliansky,et al.  Human smooth muscle VLA-1 integrin: purification, substrate specificity, localization in aorta, and expression during development , 1990, The Journal of cell biology.

[29]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[30]  Bhanu P. Jena,et al.  Journal of Cellular and Molecular Medicine , 2014, Journal of Cellular and Molecular Medicine.

[31]  S. Fowler,et al.  Nile red: a selective fluorescent stain for intracellular lipid droplets , 1985, The Journal of cell biology.

[32]  L Orci,et al.  Transforming growth factor beta stimulates collagen-matrix contraction by fibroblasts: implications for wound healing. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[33]  J. Singer,et al.  CGA-7 and HHF, two monoclonal antibodies that recognize muscle actin and react with adherent cells in human long-term bone marrow cultures. , 1985, Blood.

[34]  V. Koteliansky,et al.  Stromal cells from human long-term marrow cultures are mesenchymal cells that differentiate following a vascular smooth muscle differentiation pathway. , 1993, Blood.

[35]  S. Badylak,et al.  A perivascular origin for mesenchymal stem cells in multiple human organs. , 2008, Cell stem cell.

[36]  T. Dexter.  Stromal cell associated haemopoiesis , 1982, Journal of cellular physiology. Supplement.

[37]  D. Scadden,et al.  The haematopoietic stem cell niche at a glance , 2011, Journal of Cell Science.

[38]  Jennifer S. Park,et al.  Proteomic Profiling of Bone Marrow Mesenchymal Stem Cells upon Transforming Growth Factor β1 Stimulation* , 2004, Journal of Biological Chemistry.

[39]  M. Deschaseaux,et al.  The Hematopoietic Microenvironment: Phenotypic and Functional Characterization of Human Marrow Vascular Stromal Cells. , 1999, Hematology.

[40]  L. Humeau,et al.  Early ontogeny of the human marrow from long bones: an immunohistochemical study of hematopoiesis and its microenvironment. , 1996, Blood.

[41]  B. Lilly,et al.  Transforming Growth Factor-β (TGF-β1) Down-regulates Notch3 in Fibroblasts to Promote Smooth Muscle Gene Expression* , 2008, Journal of Biological Chemistry.

[42]  P. Bianco,et al.  Mesenchymal stem cells: revisiting history, concepts, and assays. , 2008, Cell stem cell.

[43]  K. Kirsch,et al.  Genomic organization of the melanoma-associated glycoprotein MUC18: implications for the evolution of the immunoglobulin domains. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[44]  M. Biffoni,et al.  Isolation and characterization of CD146+ multipotent mesenchymal stromal cells. , 2008, Experimental hematology.

[45]  G. Gabbiani,et al.  Alpha-smooth muscle actin is expressed in a subset of bone marrow stromal cells in normal and pathological conditions. , 1989, Virchows Archiv. B, Cell pathology including molecular pathology.

[46]  B. Sacchetti,et al.  Self-Renewing Osteoprogenitors in Bone Marrow Sinusoids Can Organize a Hematopoietic Microenvironment , 2007, Cell.

[47]  Karin Tarte,et al.  Good manufacturing practices production of mesenchymal stem/stromal cells. , 2011, Human gene therapy.