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제대혈 유래 CD34 양성세포로부터 적혈구 분화유도와 microarray를 이용한 유전자 프로파일 분석

Other Titles
 Ex Vivo Generation of RBCs from CD34+ Cells in Human Umbilical Cord Blood and Expression Profile Analysis Using Microarray 
Authors
 김창기  ;  이지혁  ;  박광일  ;  김현숙  ;  유철주  ;  라선영  ;  김현옥 
Citation
 KOREAN JOURNAL OF LABORATORY MEDICINE , Vol.25(6) : 457-464, 2005 
Journal Title
 KOREAN JOURNAL OF LABORATORY MEDICINE 
ISSN
 1598-6535 
Issue Date
2005
MeSH
Cord blood ; CD34+ cells ; Erythropoietin ; RBCs
Keywords
Cord blood ; CD34+ cells ; Erythropoietin ; RBCs
Abstract
Background :In this study, we attempted to generate RBCs from CD34+ cells in cord blood using a 3-step culture protocol and also evaluated a change in immunophenotypic characteristics and expression profile according to erythropoietin (EPO) concentrations and culture duration. Method :Using mini-MACS columns, CD34+ cells were isolated from cord blood. The culture procedure comprised three steps. For each step, cells were cultured sequentially for 7 days in a serum free liquid medium with a specific combination of growth factors for 21 days. [1st step: Flt3-ligand (Flt3-L), thrombopoietin and stem cell factor (SCF); 2nd step: IGF-1, SCF and EPO; and 3rd step: IGF-1 and EPO] To evaluate the effect of EPO on proliferation and differentiation, cells were cultured with different EPO concentrations (0, 3, 10 & 20 U/mL). Cell count and morphology were monitored during the culture. For phenotyping, antibodies to CD34, CD38, CD45 and glycophorin A (GPA) were used. The expression profile of cultured cells was analyzed by 17,000-gene microarray analysis. Result :As EPO concentration increased, cell expansion was also increased, showing a maximum expansion at 20 U/mL. The cell population showed a gradual decrease in expression of CD34 and CD45, whereas the expression of GPA was not prominent in any conditions. However, we observed increased expression in some genes associated with erythropoiesis (e.g. glycophorin A, rhesus blood group CcEe antigens). Conclusion :This study shows that erythropoietin enhances the proliferation of hematopoietic progenitor cells. Our culture system did not achieve pure production of RBCs, but induced expression changes that indicated erythroid differentiation.
Files in This Item:
T200501662.pdf Download
DOI
OAK-2005-06764
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Internal Medicine (내과학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Laboratory Medicine (진단검사의학교실) > 1. Journal Papers
5. Research Institutes (연구소) > Cancer Metastasis Research Center (암전이연구센터) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Pediatrics (소아청소년과학교실) > 1. Journal Papers
Yonsei Authors
Kim, Hyon Suk(김현숙) ORCID logo https://orcid.org/0000-0001-5662-7740
Kim, Hyun Ok(김현옥) ORCID logo https://orcid.org/0000-0002-4964-1963
Rha, Sun Young(라선영) ORCID logo https://orcid.org/0000-0002-2512-4531
Lyu, Chuhl Joo(유철주) ORCID logo https://orcid.org/0000-0001-7124-7818
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/149877
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