Skip to main content

Advertisement

SpringerLink
Log in

Role of CD9 in proliferation and proangiogenic action of human adipose-derived mesenchymal stem cells

  • Cell and Molecular Physiology
  • Published:
Pflügers Archiv - European Journal of Physiology Aims and scope Submit manuscript

Abstract

CD9 belongs to the tetraspanin family and is involved in cell motility, osteoclastogenesis, metastasis, neurite outgrowth, myotube formation, and sperm–egg fusion. CD9 also promotes juxtacrine signaling involved in proliferation and attachment. Varying degrees of CD9 expression have been found in human mesenchymal stem cells. In this study, we determined the functional roles of CD9 in human adipose-derived mesenchymal stem cells (hASCs). The CD9 expression in hASCs was down-regulated during culture expansion. A colony-forming unit assay revealed that the clonal expandability of hASCs was directly correlated with the CD9 expression level of the colony. The CD9(high) cells exhibited an increased ability to proliferate, increased cell adhesiveness, and better in vitro tube formation than the CD9(low) cells. The cellular proliferation and attachment of the CD9(high) cells were inhibited upon treatment with a blocking antibody against CD9 and the transduction of a CD9 miRNA lentivirus. The CD9(high) cells showed higher NF-κB promoter activity and higher levels of intercellular adhesion molecule 1 than the CD9(low) cells. Reverse transcription-polymerase chain reaction analysis revealed higher endothelial nitric oxide synthase expression in the CD9(high) cells than in the CD9(low) cells. The engraftment and the proangiogenic action of hASCs in a murine model of hindlimb ischemia were significantly higher in the CD9(high) cells than in the CD9(low) cells. This study indicates that CD9 plays roles in cell proliferation and attachment in vitro as well as in in vivo engraftment and that it can be considered as a useful marker to predict the in vivo efficacy of hASCs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Levy S, Shoham T (2005) Protein–protein interactions in the tetraspanin web. Physiology (Bethesda) 20:218–224

    CAS  Google Scholar 

  2. Hemler ME (2005) Tetraspanin functions and associated microdomains. Nat Rev Mol Cell Biol 6(10):801–811 (Review, Oct)

    Article  PubMed  CAS  Google Scholar 

  3. Maecker HT, Todd SC, Levy S (1997) The tetraspanin superfamily: molecular facilitators. FASEB J 11:428–442

    PubMed  CAS  Google Scholar 

  4. Boucheix C, Rubinstein E (2001) Tetraspanins. Cell Mol Life Sci 58(9):1189–1205 (Aug)

    Article  PubMed  CAS  Google Scholar 

  5. Kovalenko OV, Metcalf DG, DeGrado WF, Hemler ME (2005) Structural organization and interactions of transmembrane domains in tetraspanin proteins. BMC Struct Biol 5:11 (Jun 28)

    Article  PubMed  CAS  Google Scholar 

  6. Charrin S, Le Naour F, Oualid M, Billard M, Faure G, Hanash SM, Boucheix C, Rubinstein E (2001) The major CD9 and CD81 molecular partner. Identification and characterization of the complexes. J Biol Chem 276:14329–14337

    PubMed  CAS  Google Scholar 

  7. Boucheix C, Perrot JY, Mirshahi M (1985) A new set of monoclonal antibodies against acute lymphoblastic leukemia. Leuk Res 9:597–604

    Article  PubMed  CAS  Google Scholar 

  8. Sincock PM, Mayrhofer G, Ashman LK (1997) Localization of the transmembrane 4 superfamily (TM4SF) member PETA-3 (CD151) in normal human tissues: comparison with CD9, CD63, and alpha5beta1 integrin. J Histochem Cytochem 45(4):515–525

    PubMed  CAS  Google Scholar 

  9. Miyake M, Koyama M, Seno M (1991) Identification of the motility-related protein (MRP-1), recognized by monoclonal antibody M31-15, which inhibits cell motility. J Exp Med 174:1347–1354

    Article  PubMed  CAS  Google Scholar 

  10. Takeda Y, Tachibana I, Miyado K (2003) Tetraspanins CD9 and CD81 function to prevent the fusion of mononuclear phagocytes. J Cell Biol 16:945–956

    Article  CAS  Google Scholar 

  11. Miyake M, Nakano K, Ieki Y (1995) Motility related protein 1 (MRP-1/CD9) expression: inverse correlation with metastases in breast cancer. Cancer Res 55:4127–4131

    PubMed  CAS  Google Scholar 

  12. Miyake M, Inufusa H, Adachi M (2000) Suppression of pulmonary metastasis using adenovirally motility related protein-1 (MRP-1/CD9) gene delivery. Oncogene 19:5221–5226

    Article  PubMed  CAS  Google Scholar 

  13. Hadjiargyrou M, Patterson PH (1995) An anti-CD9 monoclonal antibody promotes adhesion and induces proliferation of Schwann cells in vitro. J Neurosci 15:574–583

    PubMed  CAS  Google Scholar 

  14. Tachibana I, Bodorova J, Berditchevski F (1997) NAG-2, a novel transmembrane-4 superfamily (TM4SF) protein that complexes with integrins and other TM4SF proteins. J Biol Chem 272:29181–29189

    Article  PubMed  CAS  Google Scholar 

  15. Chen MS, Tung KS, Coonrod SA (1999) Role of the integrin-associated protein CD9 in binding between sperm ADAM 2 and the egg integrin alpha6beta1: implications for murine fertilization. Proc Natl Acad Sci USA 96:11830–11835

    Article  PubMed  CAS  Google Scholar 

  16. Le Naour F, Rubinstein E, Jasmin C, Prenant M, Boucheix C (2000) Severely reduced female fertility in CD9-deficient mice. Science 287(5451):319–321

    Article  PubMed  Google Scholar 

  17. Miyado K, Yamada G, Yamada S, Hasuwa H, Nakamura Y, Ryu F, Suzuki K, Kosai K, Inoue K, Ogura A, Okabe M, Mekada E (2000) Requirement of CD9 on the egg plasma membrane for fertilization. Science 287(5451):321–324

    Article  PubMed  CAS  Google Scholar 

  18. Kaji K, Oda S, Shikano T, Ohnuki T, Uematsu Y, Sakagami J, Tada N, Miyazaki S, Kudo A (2000) The gamete fusion process is defective in eggs of Cd9-deficient mice. Nat Genet 24(3):279–282

    Article  PubMed  CAS  Google Scholar 

  19. Inui S, Higashiyama S, Hashimoto K (1997) Possible role of coexpression of CD9 with membrane-anchored heparin-binding EGF-like growth factor and amphiregulin in cultured human keratinocyte growth. J Cell Physiol 171:291–298

    Article  PubMed  CAS  Google Scholar 

  20. Shi W, Fan H, Shum L (2000) The tetraspanin CD9 associates with transmembrane TGF-alpha and regulates TGF-alpha-induced EGF receptor activation and cell proliferation. J Cell Biol 148:591–602

    Article  PubMed  CAS  Google Scholar 

  21. Si Z, Hersey P (1993) Expression of the neuroglandular antigen and analogues in melanoma. CD9 expression appears inversely related to metastatic potential of melanoma. Int J Cancer 54:37–43

    Article  PubMed  CAS  Google Scholar 

  22. Mori M, Mimori K, Shiraishi T (1998) Motility related protein 1 (MRP1/CD9) expression in colon cancer. Clin Cancer Res 4:1507–1510

    PubMed  CAS  Google Scholar 

  23. Mhawech P, Herrmann F, Coassin M (2003) Motility-related protein 1 (MRP-1/CD9) expression in urothelial bladder carcinoma and its relation to tumor recurrence and progression. Cancer 98:1649–1657

    Article  PubMed  CAS  Google Scholar 

  24. Higashiyama M, Taki T, Ieki Y (1995) Reduced motility related protein-1 (MRP-1/CD9) gene expression as a factor of poor prognosis in non-small cell lung cancer. Cancer Res 55:6040–6044

    PubMed  CAS  Google Scholar 

  25. Sho M, Adachi M, Taki T (1998) Transmembrane 4 superfamily as a prognostic factor in pancreatic cancer. Int J Cancer 79:509–516

    Article  PubMed  CAS  Google Scholar 

  26. Erovic BM, Pammer J, Hollemann D (2003) Motility-related protein-1/CD9 expression in head and neck squamous cell carcinoma. Head Neck 25:848–857

    Article  PubMed  Google Scholar 

  27. Mhawech P, Dulguerov P, Tschanz E (2004) Motility-related protein-1 (MRP-1/CD9) expression can predict disease-free survival in patients with squamous cell carcinoma of the head and neck. Br J Cancer 90:471–475

    Article  PubMed  CAS  Google Scholar 

  28. Miyake M, Nakano K, Itoi SI (1996) Motility-related protein-1 (MRP-1/CD9) reduction as a factor of poor prognosis in breast cancer. Cancer Res 56:1244–1249

    PubMed  CAS  Google Scholar 

  29. Murayama Y, Miyagawa J, Oritani K (2004) CD9-mediated activation of the p46 Shc isoform leads to apoptosis in cancer cells. Cell Sci 117:3379–3388

    Article  CAS  Google Scholar 

  30. Ikeyama S, Koyama M, Yamaoko M (1993) Suppression of cell motility and metastasis by transfection with human motility-related protein (MRP-1/CD9) DNA. J Exp Med 177:1231–1237

    Article  PubMed  CAS  Google Scholar 

  31. Huang CL, Liu D, Masuya D (2004) MMRP-1/CD9 gene transduction downregulates Wnt signal pathways. Oncogene 23:7475–7483

    Article  PubMed  CAS  Google Scholar 

  32. Ko EM, Lee IY, Cheon IS (2006) Monoclonal antibody to CD9 inhibits platelet-induced human endothelial cell proliferation. Mol Cells 22:70–77

    PubMed  CAS  Google Scholar 

  33. Jennings LK, Crossno JT Jr, White MM (2000) CD9 structure and function. In: Berndt MC (ed) Platelets, thrombosis and the vessel wall. Harwood Academic Publishers, Amsterdam, Holland, pp 173–187

    Google Scholar 

  34. Shaw ARE, Domanska A, Mak A (1995) Ectopic expression of human and feline CD9 in a human B cell line confers beta 1 integrin dependent motility on FN and LN substrates and enhanced tyrosine phosphorylation. J Biol Chem 270:24092–24099

    Article  PubMed  CAS  Google Scholar 

  35. Cook GA, Longhurst CM, Grgurevich S, Cholera S, Crossno JT Jr, Jennings LK (2002) Identification of CD9 extracellular domains important in regulation of CHO cell adhesion to fibronectin and fibronectin pericellular matrix assembly. Blood 100(13):4502–4511

    Article  PubMed  CAS  Google Scholar 

  36. Friedenstein AJ, Chailakhyan RK, Gerasimov UV (1987) Bone marrow osteogenic stem cells: in vitro cultivation and transplantation in diffusion chambers. Cell Tissue Kinet 20:263–272

    PubMed  CAS  Google Scholar 

  37. Friedenstein AJ, Gorskaja JF, Kulagina NN (1976) Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp Hematol 4:267–274

    PubMed  CAS  Google Scholar 

  38. Friedenstein AJ, Latzinik NW, Grosheva AG (1982) Marrow microenvironment transfer by heterotopic transplantation of freshly isolated and cultured cells in porous sponges. Exp Hematol 10:217–227

    PubMed  CAS  Google Scholar 

  39. Pittenger MF, Mackay AM, Beck SC (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147

    Article  PubMed  CAS  Google Scholar 

  40. Prockop DJ (1997) Marrow stromal cells as stem cells for nonhematopoietic tissues. Science 276:71–74

    Article  PubMed  CAS  Google Scholar 

  41. Liechty KW, MacKenzie TC, Shaaban AF (2000) Human mesenchymal stem cells engraft and demonstrate site-specific differentiation after in utero transplantation in sheep. Nature Medicine 6:1282–1286

    Article  PubMed  CAS  Google Scholar 

  42. Zuk PA, Zhu M, Mizuno H (2001) Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 7:211–228

    Article  PubMed  CAS  Google Scholar 

  43. Schaffler A, Buchler C (2007) Concise review: adipose tissue-derived stromal cells—basic and clinical implications for novel cell-based therapies. Stem Cells 25(4):818–827

    Article  PubMed  CAS  Google Scholar 

  44. Safford KM, Hicok KC, Safford SD (2002) Neurogenic differentiation of murine and human adipose-derived stromal cells. Biochem Biophys Res Commun 294:371–379

    Article  PubMed  CAS  Google Scholar 

  45. Cho HH, Bae YC, Jung JS (2006) Role of toll-like receptors on human adipose-derived stromal cells. Stem Cells 24:2744–2752

    Article  CAS  Google Scholar 

  46. Lee DH, Kang SK, Lee RH (2004) Effects of peripheral benzodiazepine receptor ligands on proliferation and differentiation of human mesenchymal stem cells. J Cell Physiol 198:91–99

    Article  PubMed  CAS  Google Scholar 

  47. Kang SK, Lee DH, Bae YC (2003) Improvement of neurological deficits by intracerebral transplantation of human adipose tissue-derived stromal cells after cerebral ischemia in rats. Exp Neurol 183:355–366

    Article  PubMed  CAS  Google Scholar 

  48. Gronthos S, Franklin DM, Leddy HA (2001) Surface protein characterization of human adipose tissue-derived stromal cells. J Cell Physiol 189:54–63

    Article  PubMed  CAS  Google Scholar 

  49. Oritani K, Aoyama K, Tomiyama Y, Kincade PW, Matsuzawa Y (2000) Stromal cell CD9 and the differentiation of hematopoietic stem/progenitor cells. Leuk Lymphoma 38(1–2):147–152 (Review)

    PubMed  CAS  Google Scholar 

  50. Moon MH, Kim SY, Kim YJ (2006) Human adipose tissue-derived mesenchymal stem cells improve postnatal neovascularization in a mouse model of hindlimb ischemia. Cell Physiol Biochem 17:279–290

    Article  PubMed  CAS  Google Scholar 

  51. Cook GA, Wilkinson WA, Crossno JT Jr (1999) The tetraspanin CD9 influences the adhesion, spreading, and pericellular FN matrix assembly of Chinese hamster ovary cells on human plasma FN. Exp Cell Res 251:356–371

    Article  PubMed  CAS  Google Scholar 

  52. Nakai K, Tanaka T, Murai T (2005) Invasive human pancreatic carcinoma cells adhere to endothelial tri-cellular corners and increase endothelial permeability. Cancer Sci 96:766–773

    Article  PubMed  CAS  Google Scholar 

  53. Brummelkamp TR, Bernards R, Agami R (2002) A system for stable expression of short interfering FNAsin mammalian cells. Science 296:550–553

    Article  PubMed  CAS  Google Scholar 

  54. Manning AM, Bell FP, Rosenbloom CL (1995) NF-kappa B is activated during acute inflammation in vivo in association with elevated endothelial cell adhesion molecule gene expression and leukocyte recruitment. J Inflam 45:283–296

    CAS  Google Scholar 

  55. Planat-Benard V, Menard C, Andre M, Puceat M, Perez A, Garcia-Verdugo JM, Penicaud L, Casteilla L (2004) Spontaneous cardiomyocyte differentiation from adipose tissue stroma cells. Circ Res 94(2):223–229

    Article  PubMed  CAS  Google Scholar 

  56. Seehafer JG, Shaw AR (1991) Evidence that the signal-initiating membrane protein CD9 is associated with small GTP-binding proteins. Biochem Biophys Res Commun 179:401–406

    Article  PubMed  CAS  Google Scholar 

  57. Kroll MH, Mendelsohn ME, Miller JL (1992) Monoclonal antibody AG-1 initiates platelet activation by a pathway dependent on glycoprotein IIb-IIIa and extracellular calcium. Biochim Biophys Acta 1137:248–256

    Article  PubMed  CAS  Google Scholar 

  58. Zilber MT, Setterblad N, Vasselon T (2005) Institut National de la Sante et de la Recherche Medicale (INSERM) U662, and Service Commun MHC class II/CD38/CD9: a lipid-raft-dependent signaling complex in human monocytes. Blood 106:3074–3081

    Article  PubMed  CAS  Google Scholar 

  59. Chung J, Mercurio AM (2004) Contributions of the alpha6 integrins to breast carcinoma survival and progression. Mol Cells 17:203–209

    PubMed  CAS  Google Scholar 

  60. Klein-Soyer C, Azorsa DO, Cazenave JP (2000) CD9 participates in endothelial cell migration during in vitro wound repair. Arterioscler Thromb Vasc Biol 20:360–369

    PubMed  CAS  Google Scholar 

  61. Slupsky JR, Cawley JC, Kaplan C (1997) Analysis of CD9, CD32 and p67 signalling: use of degranulated platelets indicates direct involvement of CD9 and p67 in integrin activation. Br J Haematol 96:275–286

    Article  PubMed  CAS  Google Scholar 

  62. Le Naour F, Andre M, Boucheix C, Rubinstein E (2006) Membrane microdomains and proteomics: lessons from tetraspanin microdomains and comparison with lipid rafts. Proteomics 6(24):6447–6454 (Review)

    Article  PubMed  CAS  Google Scholar 

  63. Le Naour F, Charrin S, Labas V, Le Caer JP, Boucheix C, Rubinstein E (2004) Tetraspanins connect several types of Ig proteins: IgM is a novel component of the tetraspanin web on B-lymphoid cells. Cancer Immunol Immunother 53(3):148–152 (Review)

    Article  PubMed  CAS  Google Scholar 

  64. Sugiura T, Berditchevski F (1999) Function of 31-tetraspanin protein complexes in tumor cell invasion. Evidence for the role of the complexes in production of matrix metalloproteinase 2 (MMP-2). J Cell Biol 146:1375–1389

    Article  PubMed  CAS  Google Scholar 

  65. Hemler ME (1998) Integrin associated proteins. Curr Opin Cell Biol 10:578–585

    Article  PubMed  CAS  Google Scholar 

  66. Little KD, Hemler ME, Stipp CS (2004) Dynamic regulation of a GPCR-tetraspanin-G protein complex on intact cells: central role of CD81 in facilitating GPR56-Galpha q/11 association. Mol Biol Cell 15(5):2375–2387

    Article  PubMed  CAS  Google Scholar 

  67. Rahman A, Anwar KN, True AL, Malik AB (1999) Thrombin-induced p65 homodimer binding to downstream NF-κB site of the promoter mediates endothelial ICAM-1 expression and neutrophil adhesion. J Immunol 162:5466–5476

    PubMed  CAS  Google Scholar 

  68. Robker RL, Collins RG, Beaudet AL, Mersmann HJ, Smith CW (2004) Leukocyte migration in adipose tissue of mice null for ICAM-1 and Mac-1 adhesion receptors. Obes Res 12(6):936–940 (Jun)

    Article  PubMed  CAS  Google Scholar 

  69. Duda DG, Fukumura D, Jain RK (2004) Role of eNOS in neovascularization: NO for endothelial progenitor cells. Trends Mol Med 10:143–145

    Article  PubMed  CAS  Google Scholar 

  70. Ying L, Hofseth LJ (2007) An emerging role for endothelial nitric oxide synthase in chronic inflammation and cancer. Cancer Res 67(4):1407–1410 (Review)

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

This work was supported by a grant (S06022HTB2580A) from the Korean Food & Drug Administration.

Author information

Authors and Affiliations

  1. Department of Physiology, School of Medicine, Pusan National University, 1 Ga, Ami-Dong, Suh-Gu, Pusan, 602-739, South Korea

    Yeon Jeong Kim, Ji Min Yu, Hye Joon Joo, Hoe Kyu Kim, Hyun Hwa Cho & Jin Sup Jung

  2. Department of Plastic Surgery, School of Medicine, Pusan National University, Pusan, 602-739, South Korea

    Yong Chan Bae

  3. Medical Research Institute, Pusan National University, Pusan, 602-739, South Korea

    Jin Sup Jung

Authors
  1. Yeon Jeong Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ji Min Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hye Joon Joo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hoe Kyu Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hyun Hwa Cho
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yong Chan Bae
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jin Sup Jung
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jin Sup Jung.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kim, Y.J., Yu, J.M., Joo, H.J. et al. Role of CD9 in proliferation and proangiogenic action of human adipose-derived mesenchymal stem cells. Pflugers Arch - Eur J Physiol 455, 283–296 (2007). https://doi.org/10.1007/s00424-007-0285-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00424-007-0285-4

Keywords

Search

Navigation