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Adherent cells generated during long-term culture of human umbilical cord blood CD34+ cells have characteristics of endothelial cells and beneficial effect on cord blood ex vivo expansion

Title
Adherent cells generated during long-term culture of human umbilical cord blood CD34+ cells have characteristics of endothelial cells and beneficial effect on cord blood ex vivo expansion
Authors
Yoo E.-S.Lee K.-E.Seo J.-W.Yoo E.-H.Lee M.-A.Im S.-A.Mun Y.-C.Lee S.-N.Huh J.-W.Kim M.-J.Jo D.-Y.Ahn J.-Y.Lee S.-M.Chung W.-S.Kim J.-H.Seong C.-M.
Ewha Authors
이순남이미애서정완정화순성주명유은선허정원이경은문영철
SCOPUS Author ID
이순남scopus; 이미애scopus; 서정완scopus; 정화순scopus; 성주명scopus; 유은선scopus; 허정원scopus; 이경은scopus; 문영철scopus
Issue Date
2003
Journal Title
Stem Cells
ISSN
1066-5099JCR Link
Citation
Stem Cells vol. 21, no. 2, pp. 228 - 235
Indexed
SCI; SCIE; SCOPUS WOS scopus
Document Type
Article
Abstract
Hematopoiesis depends on the association of hematopoietic stem cells with stromal cells that constitute the hematopoietic microenvironment. The in vitro development of the endothelial cell from umbilical cord blood (UCB) is not well established and has met very limited success. In this study, UCB CD34+ cells were cultured for 5 weeks in a stroma-free liquid culture system using thrombopoietin, flt3 ligand, and granulocyte-colony stimulating factor. By week 4-5, we found that firmly adherent fibroblast-like cells were established. These cells showed characteristics of endothelial cells expressing von Willebrand factor, human vascular cell adhesion molecule-1, human intracellular adhesion molecule-1, human CD31, E-selectin, and human macrophage. Furthermore, when comparing an ex vivo system without an established endothelial monolayer to an ex vivo system with an established endothelial monolayer, better expansion of total nucleated cells, CD34+ cells, and colony-forming units (CFUs)-granulocyte-macrophage and CFUs-granulocyteerythroid-megakaryocyte-macrophage were found during culture. This phenomenon was in part due to the fact that a significant reduction of apoptotic fractions was found in the CD34+ cells, which were cultured on the adherent monolayer for up to 5 weeks. To gather quantitative data on the number of endothelial cells derived from a given number of CD34 cells, we performed limiting dilution assay by using Poisson distribution: the number of tested cells (linear scale) producing a 37% negative culture (logarithmic scale) is the number of cells containing one endothelial cell. By this method, one endothelial cell may be found from 314 CD34+ cells after 5 weeks of culture. These results suggest that the UCB CD34+ cell fraction contains endothelial cell precursors, establishing the hematopoietic microenvironment and providing the beneficial effects through downregulating apoptosis on UCB expansion protocols. These observations may provide insight for future cellular therapy or graft engineering.
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의과대학 > 의학과 > Journal papers
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