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Endothelial progenitor cell differentiation using cryopreserved, umbilical cord blood-derived mononuclear cells.

Authors
Jang, JH; Kim, SK; Choi, JE; Kim, YJ; Lee, HW; Kang, SY; Park, JS; Choi, JH; Lim, HY; Kim, HC
Citation
Acta pharmacologica Sinica, 28(3):367-374, 2007
Journal Title
Acta pharmacologica Sinica
ISSN
1671-40831745-7254
Abstract
AIM: To investigate the endothelial differentiation potentiality of umbilical cord blood (UCB), we induced the differentiation of endothelial progenitor cells (EPC) from cryopreserved UCB-derived mononuclear cells (MNC).



METHODS: MNC from cryopreserved UCB and peripheral blood (PB) were cultured in M199 medium with endothelial cell growth supplements for 14 d. EPC were characterized by RT-PCR, flow cytometry, and immunocytochemistry analysis. The proliferation of differentiated EPC was studied by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay, and vascular endothelial growth factor (VEGF) concentration was measured using an ELISA kit. Characteristics of UCB-derived EPC were compared with those of PB-derived EPC.



RESULTS: A number of round-shaped cells were loosely attached to the bottom after 24 h culture, and numerous spindleshaped cells began to appear from the round-shaped ones on d 7. Those cells expressed endothelial markers such as, Flt-1/VEGFR-1, ecNOS, VE-cadherin, von Willebrand factor, and secreted VEGF. The patterns of endothelial markers of EPC from PB and UCB did not show striking differences. The results of the proliferation and secretion of VEGF were also similar.



CONCLUSION: We successfully cultured UCB cells stored at -196 Celsius degree into cells with the quality of endothelial cells. Those EPC could be used for angiogenic therapeutics by activating adjacent endothelial cells and enhancing angiogenesis.
MeSH terms
Biological MarkersCell Differentiation*Cells, CulturedCryopreservationEndothelial Cells/physiology*Endothelial Cells/ultrastructureFetal Blood/cytologyFetal Blood/physiology*HumansMonocytes/physiology*Stem Cells/physiology*Stem Cells/ultrastructureVascular Endothelial Growth Factor A/biosynthesisVascular Endothelial Growth Factor A/genetics
DOI
10.1111/j.1745-7254.2007.00519.x
PMID
17302999
Appears in Collections:
Journal Papers > School of Medicine / Graduate School of Medicine > Hematology-Oncology
AJOU Authors
이, 현우강, 석윤박, 준성최, 진혁김, 효철
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