말초혈액 유래 간엽전구세포의 골분화

Osteogenic Differentiation of Circulating Peripheral Blood Derived Mesenchymal Progenitor Cells

  • 은석찬 (서울대학교 의과대학 성형외과학교실) ;
  • 김진희 (서울대학교 의과대학 성형외과학교실) ;
  • 허찬영 (서울대학교 의과대학 성형외과학교실) ;
  • 백롱민 (서울대학교 의과대학 성형외과학교실) ;
  • 장학 (서울대학교 의과대학 성형외과학교실) ;
  • 민경원 (서울대학교 의과대학 성형외과학교실)
  • Eun, Seok Chan (Department of Plastic and Reconstructive Surgery, Seoul National University College of Medicine) ;
  • Kim, Jin Hee (Department of Plastic and Reconstructive Surgery, Seoul National University College of Medicine) ;
  • Heo, Chan Yeong (Department of Plastic and Reconstructive Surgery, Seoul National University College of Medicine) ;
  • Baek, Rong Min (Department of Plastic and Reconstructive Surgery, Seoul National University College of Medicine) ;
  • Chang, Hak (Department of Plastic and Reconstructive Surgery, Seoul National University College of Medicine) ;
  • Minn, Kyung Won (Department of Plastic and Reconstructive Surgery, Seoul National University College of Medicine)
  • 투고 : 2008.01.21
  • 발행 : 2008.05.10

초록

Purpose: There are some reports presenting that peripheral blood contain circulating hematopoietic cells as well as, in significantly smaller quantities, mesenchymal stem cells. The purposes of this study is to isolate and characterize circulating mesenchymal progenitor cells with osteogenic potential from human peripheral blood. Methods: Human buffycoat containing mononuclear cells was harvested from peripheral blood of normal persons and isolated using a density gradient centrifugation and serially subcultured in osteogenic media for 1-4 weeks. The proliferation capability, phase-contrast microscopy, transmission electron microscopy, immunophenotype FACS analysis, Alizarin red staining and RT-PCR assays for osteogenic differentiation potential were performed. Results: The phenotype of cultured cells changed from small round or cuboidal cells at passage 1 into large spindle-shaped fibroblastic morphology cells at passage 4. Surface marker expressed CD14, but did not express CD34, CD80, CD83. Strong positive staining was observed for Alizarin reds in osteogenic medium on day 14, Using RT-PCR, the mRNA levels of bone- specific genes, such as ALP, c-bfa-1 and osteocalcin were detected. Conclusion: A new subset of peripheral blood derived progenitor cells described here has the ability to proliferate and differentiate into osteogenic cell lineages in vitro, and to be candidate for regenerative therapy.

키워드

과제정보

연구 과제 주관 기관 : 한국과학재단

참고문헌

  1. Jung EJ, Yoon TR, Song EK, Kim SH: In vitro and in vivo osteogenic induction of mesenchymal stem cells from rabbit bone marrow. J Korean Orthop Res Soc 4: 79, 2001
  2. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR: Multilineage potential of adult human mesenchymal stem cells. Science 284: 143, 1999 https://doi.org/10.1126/science.284.5411.143
  3. Oh IH, Kim DW: Three-dimensional approach to stem cell therapy. J Korean Med Sci 17: 151, 2002 https://doi.org/10.3346/jkms.2002.17.2.151
  4. Bonde J, Hess DA, Nolta JA: Recent advances in hematopoietic stem cell biology. Curr Opin Hematol 11: 392, 2004 https://doi.org/10.1097/01.moh.0000145672.42503.70
  5. Zhao Y, Glesne D, Huberman E: A human peripheral blood monocyte-derived subset acts as pluripotent stem cells. Proc Natl Acad Sci U S A 100: 2426, 2003
  6. Zvaifler NJ, Marinova-Mutafchieva L, Adams G, Edwards CJ, Moss J, Burger JA, Maini RN: Mesenchymal precursor cells in the blood of normal individuals. Arthritis Res 2: 477, 2000 https://doi.org/10.1186/ar130
  7. Hong SJ, Lee EA, Chae GT, Han H: Differentiation of osteoblast progenitor cells from human umbilical cord blood. Immune Netw 2: 166, 2002 https://doi.org/10.4110/in.2002.2.3.166
  8. Li WJ, Tuli R, Huang X, Laquerriere P, Tuan RS: Multilineage differentiation of human mesenchymal stem cells in a three-dimensional nanofibrous scaffold. Biomaterials 26: 5158, 2005 https://doi.org/10.1016/j.biomaterials.2005.01.002
  9. Spangrude GJ: When is a stem cell really a stem cell? Bone Marrow Transplant 32: S7, 2003 https://doi.org/10.1038/sj.bmt.1703936
  10. Ogawa BM: Differentiation and proliferation of hematopoietic stem cells. Blood 81: 2844, 1993
  11. Chim H, Schantz JT: Human circulating peripheral blood mononuclear cells for calvarial bone tissue engineering. Plast Reconstr Surg 117: 468, 2006 https://doi.org/10.1097/01.prs.0000201489.65811.e7
  12. Fernandez M, Simon V, Herrera G, Cao C, Del Favero H, Minguell JJ: Detection of stromal cells in peripheral blood progenitor cell collections from breast cancer patients. Bone Marrow Transplant 20: 265, 1997 https://doi.org/10.1038/sj.bmt.1700890
  13. Simper D, Stalboerger PG, Panetta CJ, Wang S, Caplice NM: Smooth muscle progenitor cells in human blood. Circulation 106: 1199, 2002 https://doi.org/10.1161/01.CIR.0000031525.61826.A8
  14. Kim SJ, Kim KH, Jung JS, Choi OH: Study on the Culture and in vivo transplantation of mesenchymal stem cells derived from human umbilical cord blood. Korean J Obstet Gynecol 48: 1402, 2005
  15. Kuznetsov SA, Mankani MH, Gronthos S: Circulation skeletal stem cells. J Cell Biol 153: 1133, 2001 https://doi.org/10.1083/jcb.153.5.1133