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Changes in the expression of CD106, osteogenic genes, and transcription factors involved in the osteogenic differentiation of human bone marrow mesenchymal stem cells

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Abstract

Mesenchymal stem cells (MSCs) are well known to possess multipotential differentiation and are becoming a good tool for clinical research. However, specific markers for their purification and the mechanism of their osteogenic differentiation remain to be elucidated. In the present study, we compared the expression of CD106, and osteogenic differentiation-related proteins and genes in human bone marrow (BM)-derived MSCs, before and after differentiation by FACS, histochemical staining, immunohistochemical staining, RT-PCR, and real-time PCR. It was found that MSCs were positive for CD13, CD29, CD44, CD73, CD90, CD105, and CD166, but negative for CD14, CD31, CD34, CD62E, CD45, and GlyA. Notably, CD106 was detected before osteogenic induction, but its expression was downregulated 10 fold after 2 weeks of osteogenic differentiation as determined by flow cytometry. The results of RT-PCR and real-time PCR revealed that the expression of CD106 mRNA in MSCs significantly decreased by 7.1-, 4.2-, and 5.1-fold, respectively after osteogenic, chondrogenic, and adipogenic differentiation. In contrast, other MSC-positive markers described above did not change significantly even after differentiation. Compared to levels in control cells, after 2 weeks of osteogenic differentiation, mRNA levels of alkaline phosphatase, bone sialoprotein, osteocalcin, and transcript factors RUNX2 and Osterix showed more than 2-fold, 5-fold, 1.5-fold, 2-fold, and 5-fold increase, respectively. Thus, we speculate that CD106 might be a useful surface marker for BMMSCs. Moreover, alkaline phosphatase, type I collagen, osteonectin, osteopontin, and biglycin were expressed in the early stages of osteogenic differentiation before bone sialoprotein and osteocalcin. The present study should help to provide a novel marker for isolating purified MSCs and characterizing osteogenic differentiation.

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References

  1. Fridenshtein A (1982) Stromal bone marrow cells and the hematopoietic microenvironment. Arkh Patol 44:3–11

    PubMed  Google Scholar 

  2. Reyes M, Lund T, Lenvik T, Aguiar D, Koodie L, Verfaillie CM (2001) Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells. Blood 98:2615–2625

    Article  PubMed  CAS  Google Scholar 

  3. Hung SC, Chen NJ, Hsieh SL, Li H, Ma HL, Lo WH (2002) Isolation and characterization of size-sieved stem cells from human bone marrow. Stem Cells 20:249–258

    Article  PubMed  Google Scholar 

  4. Sabatini F, Petecchia L, Tavian M, Villeroche VJ, Rossi GA, Brouty-Boye D (2005) Human bronchial fibroblasts exhibit a mesenchymal stem cell phenotype and multilineage differentiating potentialities. Lab Invest 30:1–10

    Google Scholar 

  5. Horwitz EM, Gordon PL, Koo WK, Marx JC, MD, Muul L, Hofmann T (2002) Isolated allogeneic bone marrow-derived mesenchymal cells engraft and stimulate growth in children with osteogenesis imperfecta: implications for cell therapy of bone. Proc Natl Acad Sci USA 99:8932–8937

    Article  PubMed  CAS  Google Scholar 

  6. Lecanda F, Avioli LV, Cheng SL (1997) Regulation of bone matrix protein expression and induction of differentiation of human osteoblasts and human bone marrow stromal cells by bone morphogenetic protein-2. J Cell Biochem 67:386–396

    Article  PubMed  CAS  Google Scholar 

  7. Stewart K, Monk P, Walsh S, Jefferiss CM, Letchford J, Beresford JN (2003) STRO-1, HOP-26 (CD63), CD49a, and SB-10 (CD166) as markers of primitive human marrow stromal cells and their more differentiated progeny: a comparative investigation in vitro. Cell Tissue Res 313:281–290

    Article  PubMed  CAS  Google Scholar 

  8. Oswald J, Jorgensen SB, Feldmann S, Ehninger G, Bornhauser M, Werner C (2004) Mesenchymal stem cells can be differentiated into endothelial cells in vitro. Stem Cells 22:377–384

    Article  PubMed  Google Scholar 

  9. Shahdadfar A, Frønsdal K, Haug T, Reinholt FP, Brinchmann JE (2005) In vitro expansion of human mesenchymal stem cells: choice of serum is a determinant of cell proliferation, differentiation, gene expression and transcriptome stability. Stem Cells 23:1357–1366

    Article  PubMed  CAS  Google Scholar 

  10. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147

    Article  PubMed  CAS  Google Scholar 

  11. Ying QL, Nichols J, Chambers I, Smith A (2003) BMP induction of Id proteins suppresses differentiation and sustains embryonic stem cell self-renewal in collaboration with STAT3. Cell 115: 281–292

    Article  PubMed  CAS  Google Scholar 

  12. Qi H, Aguiar DJ, Williams SM, Pean AL, Pan W, Verfaillie CM (2003) Identification of genes responsible for osteoblast differentiation from human mesodermal progenitor cells. Proc Natl Acad Sci U S A 100:3305–3310

    Article  PubMed  CAS  Google Scholar 

  13. Solchaga LA, Penick K, Porter JD, Goldberg VM, Caplan AI, Welter JF (2005) FGF-2 enhances the mitotic and chondrogenic potentials of human adult bone marrow-derived mesenchymal stem cells. J Cell Physiol 203:398–409

    Article  PubMed  CAS  Google Scholar 

  14. Ikeda R, Yoshida K, Tsukahara S, Sakamoto Y, Tanaka H, Furukawa K, Inoue I (2005) The promyelotic leukemia zinc finger promotes osteoblastic differentiation of human mesenchymal stem cells as an upstream regulator of CBFA1. J Biol Chem 280:8523–8530

    Article  PubMed  CAS  Google Scholar 

  15. Friedman MS, Long MW, Hankenson KD (2006) Osteogenic differentiation of human mesenchymal stem cells is regulated by bone morphogenetic protein-6. J Cell Biochem 98:538–554

    Article  PubMed  CAS  Google Scholar 

  16. Franceschi RT (2005) Biological approaches to bone regeneration by gene therapy. J Dent Res 84:1093–1103

    Article  PubMed  CAS  Google Scholar 

  17. Viereck V, Siggelkow H, Tauber S, Raddatz D, Schutze N, Hufner M (2002) Differential regulation of Cbfa1/Runx2 and osteocalcin gene expression by vitamin-D3, dexamethasone, and local growth factors in primary human osteoblasts. J Cell Biochem 86:348–356

    Article  PubMed  CAS  Google Scholar 

  18. Chaudhary LR, Hofmeister AM, Hruska KA (2004) Differential growth factor control of bone formation through osteoprogenitor differentiation. Bone (NY) 34:402–411

    CAS  Google Scholar 

  19. Akiyama Y, Kuzushima K, Tsurumi T, Yamaguchi K (2004) Analysis of HLA-A24-restricted CMVpp65 peptide-specific CTL with HLA-A*2402-CMVpp65 tetramer. Immunol Lett 95:199–205

    Article  PubMed  CAS  Google Scholar 

  20. Akiyama Y, Maruyama K, Nara N, Mochizuki T, Yamamoto A, Amazaki NY, Kawashima I, Nukaya I, Takesako K, Yamaguchi K (2004) Cytotoxic T cell induction against human malignant melanoma cells using HLA-A24-restricted melanoma peptide cocktail. Anticancer Res 24:571–578

    PubMed  CAS  Google Scholar 

  21. Karlsson C, Brantsing C, Svensson T, Brisby H, Asp J, Tallheden T, Lindahl A (2007) Differentiation of human mesenchymal stem cells and articular chondrocytes: analysis of chondrogenic potential and expression pattern of differentiation-related transcription factors. J Orthop Res 25:152–163

    Article  PubMed  CAS  Google Scholar 

  22. Mayer H, Bertram H, Lindenmaier W, Korff T, Weber H, Weich H (2005) Vascular endothelial growth factor (VEGF-A) expression in human mesenchymal stem cells: autocrine and paracrine role on osteoblastic and endothelial differentiation. J Cell Biocheml 95:827–839

    Article  CAS  Google Scholar 

  23. Madras N, Gibbs AL, Zhou Y, Zandstra PW, Aubin JE (2002) Modeling stem cell development by retrospective analysis of gene expression profiles in single progenitor-derived colonies. Stem Cells 20:230–240

    Article  PubMed  CAS  Google Scholar 

  24. Shur I, Lokiec F, Bleiberg I, Benayahu D (2001) Differential gene expression of cultured human osteoblasts. J Cell Biochem 83:547–553

    Article  PubMed  CAS  Google Scholar 

  25. Maurer CA, Friess H, Kretschmann B, Wildi S, Muller C, Graber H, Schilling M, Buchler MW (1998) Over-expression of ICAM-1, VCAM-1 and ELAM-1 might influence tumor progression in colorectal cancer. Int J Cancer 79:76–81

    Article  PubMed  CAS  Google Scholar 

  26. Ali S, Kaur J, Patel KD (2000) Intercellular cell adhesion molecule-1, vascular cell adhesion molecule-1, and regulated on activation normal T cell expressed and secreted are expressed by human breast carcinoma cells and support eosinophil adhesion and activation. Am J Pathol 157:313–321

    PubMed  CAS  Google Scholar 

  27. Banks RE, Gearing AJ, Hemingway IK, Norfolk DR, Perren TJ, Selby PJ (1993) Circulating intercellular molecule-1 (ICAM-1), E-selectin and vascular cell adhesion molecule-1 (VCAM-1) in human malignancies. Br J Cancer 68:122–124

    PubMed  CAS  Google Scholar 

  28. Djouad F, PLence P, Bony C, Tropel P, Apparailly F, Sany J, Noel D, Jorgensen C (2003) Immunosuppressive effect of mesenchymal stem cells favors tumor growth in allogeneic animals. Blood 102:3837–3844

    Article  PubMed  CAS  Google Scholar 

  29. Lin KY, Lu D, Hung CF, Peng S, Huang L, Jie C, Murilo F, Rowley J, Tsai YC, He L, Kim DJ, Jaffee E, Pardoll D, Wu TC (2007) Ectopic expression of vascular cell adhesion molecule-1 as a new mechanism for tumor immune evasion. Cancer Res 67:1832–1841

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Immunotherapy Division, Shizuoka Cancer Center Research Institute, 1007 Shimonagakubo, Nagaizumi-cho, Sunto-gun, Shizuoka, 411-8777, Japan

    Feng Liu, Yasuto Akiyama, Sachiko Tai, Kouji Maruyama, Yoshihiro Kawaguchi & Ken Yamaguchi

  2. Cancer Diagnostics Research Division, Shizuoka Cancer Center Research Institute, Shizuoka, Japan

    Kouji Muramatsu

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  1. Feng Liu
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  2. Yasuto Akiyama
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  3. Sachiko Tai
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  4. Kouji Maruyama
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  5. Yoshihiro Kawaguchi
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  6. Kouji Muramatsu
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  7. Ken Yamaguchi
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Correspondence to Yasuto Akiyama.

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Electronic supplementary material The online version of this article (doi: DOI 10.1007/s00774-007-0842-0) contains supplementary material, which is available to authorized users.

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Liu, F., Akiyama, Y., Tai, S. et al. Changes in the expression of CD106, osteogenic genes, and transcription factors involved in the osteogenic differentiation of human bone marrow mesenchymal stem cells. J Bone Miner Metab 26, 312–320 (2008). https://doi.org/10.1007/s00774-007-0842-0

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  • DOI: https://doi.org/10.1007/s00774-007-0842-0

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