Osteogenic Differentiation of Circulating Peripheral Blood Derived Mesenchymal Progenitor Cells.

Eun, Seok Chan; Kim, Jin Hee; Heo, Chan Yeong; Baek, Rong Min; Chang, Hak; Minn, Kyung Won

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Original Article

Journal of the Korean Society of Plastic and Reconstructive Surgeons 2008;35(3):229-234.
Published online May 1, 2008.

Osteogenic Differentiation of Circulating Peripheral Blood Derived Mesenchymal Progenitor Cells.

Seok Chan Eun, Jin Hee Kim, Chan Yeong Heo, Rong Min Baek, Hak Chang, Kyung Won Minn

Department of Plastic and Reconstructive Surgery, Seoul National University College of Medicine, Korea. sceun@snubh.org

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: Peripheral blood; Mesenchymal Progenitor cell; Osteogenesis

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