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Amirizadeh N, Zaker F, Nasiri N. Evaluation of Umbilical Cord Blood CD34+ Hematopoietic Stem Cell Expansion in Co-culture with BM Mesenchymal Stem Cells in Presence of TEPA. Sci J Iran Blood Transfus Organ 2014; 10 (4) :353-363
URL: http://bloodjournal.ir/article-1-827-en.html
Evaluation of Umbilical Cord Blood CD34+ Hematopoietic Stem Cell Expansion in Co-culture with BM Mesenchymal Stem Cells in Presence of TEPA
N. Amirizadeh , F. Zaker , N. Nasiri *
Keywords: Key words: Hematopoietic Stem Cells, Mesenchymal Stromal Cells, TEPA
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Original Article
 
 
 
 
Sci J Iran Blood Transfus Organ 2014; 10(4): 353-363
 
 
 

Evaluation of Umbilical Cord Blood CD34+ Hematopoietic
 Stem Cell Expansion in Co-culture with BM Mesenchymal
Stem Cells in Presence of TEPA
 
Amirizadeh N.1, Zaker F.2, Nasiri N.3
 
1Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran
2School of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran
3Tehran University of Medical Sciences, Tehran, Iran
 
Abstract
Background and Objectives
Hematopoietic stem cells (HSCs) have become a standard protocol for the treatment of many hematologic malignancies and non-malignant disorders. Umbilical cord blood, as a source of HSCs, has many advantages compared to other sources. One major drawback in using this source in transplantation is the low HSC dose available. Ex vivo expansion of HSCs is a solution to overcome this limitation .In this study, we used TEPA, as a Cu chelator, and human bone marrow MSCs to investigate the expansion rate of UCB-HSCs.
 
Materials and Methods
CB-HSCs were isolated using miniMACS magnetic separation system. We cultured the enriched CD34+ cells in various conditions. Culture condition A, supplemented only with recombinant cytokines; culture condition B, supplemented with BM-MSCs as a cell feeder layer and recombinant cytokines; culture condition C, supplemented with recombinant cytokines and TEPA; culture condition D, supplemented with recombinant cytokines, BM-MSCs as a cell feeder layer and TEPA. In order to evaluate the HSC expansion, we performed cell count, the analysis of CD34+ expression by flow cytometery, and CFU assay on day 10 after culture.
 
Results
The most fold increase rate in CD34+ cell, TNCs and CFU-C was observed in the culture condition D (110.11 ± 15.3, 118.5 ± 21 and 172.9 ± 44.7, respectively) compared to other conditions.
 
Conclusions 
The results showed that co-culture of HSCs with BM-MSCs in the presence of copper chelating agent (TEPA) could dramatically increase the expansion rate of UCB-HSCs. Therefore, this strategy could be useful for HSC expansion.
 
Key words: Hematopoietic Stem Cells, Mesenchymal Stromal Cells, TEPA
 
 
 
Received: 23 Jan 2012
Accepted:10 Apr 2013
 
 
 

Correspondence: Nasiri N., MSc of Hematology. School of Allied Medical Sciences, Tehran University of Medical Sciences.
P.O.Box: 14155-6183, Tehran, Iran. Tel: (+9821) 88622576; Fax: (+9821) 88622576
E-mail: nahid.nasiri64@yahoo.com
Type of Study: Research | Subject: Stem cells
Published: 2013/12/18
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