Osteogenic Differentiation of Circulating Peripheral Blood Derived Mesenchymal Progenitor Cells

말초혈액 유래 간엽전구세포의 골분화

  • 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)
  • 은석찬 (서울대학교 의과대학 성형외과학교실) ;
  • 김진희 (서울대학교 의과대학 성형외과학교실) ;
  • 허찬영 (서울대학교 의과대학 성형외과학교실) ;
  • 백롱민 (서울대학교 의과대학 성형외과학교실) ;
  • 장학 (서울대학교 의과대학 성형외과학교실) ;
  • 민경원 (서울대학교 의과대학 성형외과학교실)
  • Received : 2008.01.21
  • Published : 2008.05.10

Abstract

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.

Keywords

Acknowledgement

Supported by : 한국과학재단

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