View : 56 Download: 0

Ultrastructural and phenotypic analysis of in vitro erythropoiesis from human cord blood CD34+ cells

Ultrastructural and phenotypic analysis of in vitro erythropoiesis from human cord blood CD34+ cells
Kie J.-H.Jung Y.-J.Woo S.-Y.Ryu K.-H.Park H.-Y.Chung W.-S.Seoh J.-Y.
Ewha Authors
서주영scopus; 정화순scopus; 유경하scopus; 우소연scopus
Issue Date
Journal Title
Annals of Hematology
0939-5555JCR Link
Annals of Hematology vol. 82, no. 5, pp. 278 - 283
Document Type
Erythropoietin (EPO) induces erythropoiesis in vitro as well as in vivo, and the process of erythroid differentiation has been explored phenotypically and morphologically. However, morphological analysis of in vitro erythropoiesis of human hematopoietic progenitor cells at the ultrastructural level has not been reported before. In the present study, we have traced the ultrastructural changes of erythroid differentiation during ex vivo expansion of human cord blood (CB) CD34+ cells in the presence of EPO by electron microscopy (EM), along with concurrent phenotypic analysis. CD34+ cells purified from ten CBs by immunomagnetic selection were cultured in serum-free essential media in the presence of a combination of the several cytokines including EPO, thrombopoietin, flt3-ligand (FL), stem cell factor (SCF), granulocyte colony-stimulating factor, interleukin (IL)-3 and/or IL-11. Phenotypic analysis was performed by flow cytometric analysis for erythroid markers, including glycophorin C (GPC), Kell-related, glycophorin A (GPA , band 3, Lub , and RhD. Ultrastructural analysis was performed by electron-microscopic examination of the cultured cells stained with uranyl acetate and lead citrate. Phenotypic analysis revealed that in the absence of EPO, genuine erythroid fraction expressing the typical pattern of erythroid markers did not appear. The order of the above markers expressed in the cultured cells in the presence of EPO was GPC, Kell-related, GPA, band 3, Lub, and RhD, irrespective of the type of cytokine added. Of the cytokines used in combination with EPO, FL + IL-3 was the most efficient in inducing erythroid differentiation, which was followed by SCF + IL-3. EM examination demonstrated complete process of erythroid development from pronormoblasts to reticulocytes with nuclei having been extruded and mature erythrocytes. These results suggest that morphologically intact erythrocytes could be produced by ex vivo expansion of CB CD34+ cells using EPO.
Appears in Collections:
의과대학 > 의학과 > Journal papers
Files in This Item:
There are no files associated with this item.
RIS (EndNote)
XLS (Excel)


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.