Isolation of Endothelial Progenitor Cells from Cord Blood and Induction of Differentiation by Ex Vivo Expansion.
10.3349/ymj.2005.46.2.260
- Author:
Jeong Won SHIN
1
;
Dong Wha LEE
;
Moon Jung KIM
;
Kyung Soon SONG
;
Han Soo KIM
;
Hyun Ok KIM
Author Information
1. Department of Laboratory Medicine, Soonchunhyang University Hospital, Korea.
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
Endothellial progenitor cells;
neovascularization;
thrombopoietin
- MeSH:
Antigens, CD34/analysis;
Cell Differentiation/drug effects;
Cell Division/drug effects;
*Cell Separation;
Cells, Cultured;
Cytokines/pharmacology;
*Endothelial Cells/immunology;
Fetal Blood/*cytology;
Fetus;
Flow Cytometry;
Humans;
Research Support, Non-U.S. Gov't;
*Stem Cells/*immunology;
Thrombopoietin/pharmacology
- From:Yonsei Medical Journal
2005;46(2):260-267
- CountryRepublic of Korea
- Language:English
-
Abstract:
Endothelial progenitor cells (EPCs) have been reported to possess the capacity to colonize vascular grafts and hold promise for therapeutic neovascularization. However, limited quantities of EPCs have been the major factor impeding effective research on vasculoangiogenesis. In this study, cytokine and culture conditions necessary for the provision of large quantities of endothelial cells (ECs) were investigated. Cord blood was collected from 18 normal full-term deliveries and CD34+ cells were isolated by MACS system (Miltenyi Biotech, Bergish-Gladbach, Germany). To evaluate the effect of cytokines, CD34+ cells were cultured with various cytokine combinations, such as stem cell factor (SCF), flt3-ligand (FL), and thrombopoietin (TPO) with vascular endothelial growth factor (VEGF), interleukin-1beta, fibroblast growth factor-basic (FGF-b) as basic cytokines. The quantities of non-adherent and adherent cells were the greatest with SCF, FL and TPO. The addition of TPO to all other cytokines significantly increased the number of non-adherent and adherent cells (p< 0.05, Wilcoxon rank sum test). After four weeks of culture, adherent cells expressed endothelial specific markers such as KDR, CD31 and CD62E. Typical morphology of ECs was observed during culture, such as cord-like structure and cobblestone appearance, suggesting that the adherent cells were consistent with ECs. In this study, the experimental conditions that optimize the production of ECs for therapeutic neovascularization were described. And it was possibly suggested that TPO plays a major role in differentiation from EPCs to ECs.
