J Korean Soc Plast Reconstr Surg.  2008 May;35(3):229-234.

Osteogenic Differentiation of Circulating Peripheral Blood Derived Mesenchymal Progenitor Cells

• Eun SC
• Kim JH
• Heo CY
• Baek RM
• Chang H
• Minn KW

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.

Keyword

Peripheral blood; Mesenchymal Progenitor cell; Osteogenesis

MeSH Terms

Anthraquinones
Cell Lineage
Cells, Cultured
Centrifugation, Density Gradient
Durapatite
Fibroblasts
Humans
Mesenchymal Stromal Cells
Microscopy, Electron, Transmission
Microscopy, Phase-Contrast
Osteocalcin
Osteogenesis
Phenotype
Polymerase Chain Reaction
RNA, Messenger
Stem Cells
Anthraquinones
Durapatite
Osteocalcin
RNA, Messenger