Volume 14, Issue 2 (2011)
mjms 2011, 14(2): 1-12 |
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Atashi A, Kaviani S, Soleimani M, Norouzinia M, Mortazavi Y, Hafizi M. Differentiation of cord blood stem cells into erythroid progenitor cells in semisolid culture media containing SCF, IL-3, IL-6 and EPO. mjms 2011; 14 (2) :1-12
URL: http://mjms.modares.ac.ir/article-30-4184-en.html
URL: http://mjms.modares.ac.ir/article-30-4184-en.html
Amir Atashi1 
, Saeid Kaviani 1, Masoud Soleimani , Mehrdad Norouzinia2 , Yousef Mortazavi3 , Maryam Hafizi4
, Saeid Kaviani 1, Masoud Soleimani , Mehrdad Norouzinia2 , Yousef Mortazavi3 , Maryam Hafizi4
1- Department of Hematology and Blood Banking, Faculty of Medical Sciences, Tarbiat Modares University
2- Department of Medical Genetic, Faculty of Medical Sciences, Tarbiat Modares University
3- Associated Professor, Department of Pathology, Faculty of Medicine, Zanjan University of Medical Sciences
4- Department of Stem Cell Biology, Stem Cell Technology Research center
2- Department of Medical Genetic, Faculty of Medical Sciences, Tarbiat Modares University
3- Associated Professor, Department of Pathology, Faculty of Medicine, Zanjan University of Medical Sciences
4- Department of Stem Cell Biology, Stem Cell Technology Research center
Abstract: (5859 Views)
Objectives: The study of erythropoiesis needs to develop the methods for erythroid progenitor cells (EPCs) culture using stem cell potency to differentiate into variety of hematopoietic cells lineages. In this study, we induced differentiation of cord blood stem cells into erythroid progenitor cells in a semisolid culture media.
Materials and Methods: Cord blood mononuclear cells were isolated and cultured in liquid culture media consist of erythroid differentiation factors (phase I). Non-adherent cells were cultured in semisolid media containing SCF, IL-3, IL-6 and EPO (phase II). After one week, the appeared erythroid colonies were picked up. Characterization of differentiated cells was performed by assessment of CD235a and CD36 using flowcytometry, giemsa cytological staining and gene expression analysis of GATA-1, EKLF, α-globin genes by RT-PCR.
Results: Flowcytometry analysis to detect CD235a and CD36 positive cells showed that 94.3% and 95.5% of differentiated cells have erythroid specific cell markers, respectively. Morphology of the cells in giemsa stained slides demonstrated erythroid progenitor cells, ranged from proerythroblast to orthochromatic erythroblast. The RT-PCR results, confirmed the precursor state of erythroid differentiated cells by expression of GATA-1, EKLF, α-globin genes.
Conclusions: Cord blood stem cells have high potency to differentiate into erythroid progenitor cells using described method that can be utilized in the experimental studies.
Article Type: Original Manuscipt |
Subject:
Medical Biotechnology|Hematology
Received: 2010/12/27 | Accepted: 2011/08/27
Received: 2010/12/27 | Accepted: 2011/08/27
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