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:: Volume 12, Issue 3 (Autumn 2015) ::
Sci J Iran Blood Transfus Organ 2015, 12(3): 244-254 Back to browse issues page
Retinal cells development through transfection of CD133+ HSC derived from human cord blood by PAX6 master gene
S. Balagholi , S. Amini-Kafi Abad , Z.S. Soheili , Sh. Samiee , H. Ahmadiyeh , M. Rezaei Kanavi , M. Nikougoftar Zarif , N. Daftarian
Abstract:   (7326 Views)

  Abstract

 Background and Objectives

 Cord blood CD133+ stem cells are highly proliferative primitive hematopoietic progenitors. These cells can also differentiate to other cells especially neural cells. The PAX6 gene belongs to a family of genes that play a critical role in the formation of tissues and organs during embryonic development. The PAX6 protein is thought to activate genes involved in the formation of the eyes, brain and spinal cord (central nervous system), and the pancreas. The purpose of this study was to assess PAX6 (5a) gene transfer in cord blood CD133+ stem cells and evaluate its effect on differentiation.

 

 Materials and Methods

 In this experimental study, cord blood stem cells were collected and mononuclear cells isolated by ficoll then, CD133+ cells were obtained using the CD133 MicroBead Kit in combination with the autoMACS Separator. These cells were cultured in Stem Span media. Next, HEK293T packaging cells were co-transfected with PLEX-MCS, PsPAX2, and pMD2G by calcium phosphate method. Lentiviral vectors were collected and concentrated. The appropriate amount of viruses was used to infect CD133+ cells. The successful transduced cells were selected by puromycin resistance. After two weeks, the expression of Rhodopsin, CHX10, Thy1, Nestin, and PAX6 proteins was assessed by immunocytochemisrty method.

  

 Results

 Two weeks after infection, the expression of Rhodopsin, CHX10, Thy1, Nestin, and PAX6 proteins were detected in treatment cells and the expression of Rhodopsin and Nestin in control cells.

  

 Conclusions

 The results showed CD133+ cells differentiated into progenitor neural like cells and retinal neural like cells including ganglion like cells.

 

Keywords: Key words: PAX6 protein, CD133 antigen, Cord Blood
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Type of Study: Research | Subject: Hematology
Published: 2015/09/2
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Balagholi S, Amini-Kafi Abad S, Soheili Z, Samiee S, Ahmadiyeh H, Rezaei Kanavi M, et al . Retinal cells development through transfection of CD133+ HSC derived from human cord blood by PAX6 master gene. Sci J Iran Blood Transfus Organ 2015; 12 (3) :244-254
URL: http://bloodjournal.ir/article-1-861-en.html


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Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Volume 12, Issue 3 (Autumn 2015) Back to browse issues page
فصلنامه پژوهشی خون Scientific Journal of Iran Blood Transfus Organ
The Scientific Journal of Iranian Blood Transfusion Organization - Copyright 2006 by IBTO
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