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miR-146a and miR-150 promote the differentiation of CD133+ cells into T-lymphoid lineage
Authors:Parviz Fallah  Ehsan Arefian  Mahmood Naderi  Seyed Hamid Aghaee-Bakhtiari  Amir Atashi  Katayoun Ahmadi  Abbas Shafiee  Masoud Soleimani
Institution:1. Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, 1997775555, Tehran, Iran
2. Department of Pathobiology, School of Medicine, Alborz University of Medical Sciences, Karaj, Iran
3. Microbial Biotechnology Laboratory, Department of Microbiology, School of Biology, College of Science, University of Tehran, 14155-6455, Tehran, Iran
4. Department of Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
5. Department of Hematology, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran
6. Department of Tissue Engineering, School of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran, Iran
Abstract:MicroRNAs control the genes involved in hematopoietic stem cell (HSCs) survival, proliferation and differentiation. The over-expression of miR-146 and miR-150 has been reported during differentiation of HSCs into T-lymphoid lineage. Therefore, in this study we evaluated the effect of their over-expression on CD133+ cells differentiation to T cells. miR-146a and miR-150 were separately and jointly transduced to human cord blood derived CD133+ cells (>97 % purity). We used qRT-PCR to assess the expression of CD2, CD3ε, CD4, CD8, CD25, T cell receptor alpha (TCR-α) and Ikaros genes in differentiated cells 4 and 8 days after transduction of the miRNAs. Following the over-expression of miR-146a, significant up-regulation of CD2, CD4, CD25 and Ikaros genes were observed (P < 0.01). On the other hand, over-expression of miR-150 caused an increase in the expression of Ikaros, CD4, CD25 and TCR-α. To evaluate the combinatorial effect of miR-146a and miR-150, transduction of both miRNAs was concurrently performed which led to increase in the expression of Ikaros, CD4 and CD3 genes. In conclusion, it seems that the effect of miR-150 and miR-146a on the promotion of T cell differentiation is time-dependant. Moreover, miRNAs could be used either as substitutes or complements of the conventional differentiation protocols for higher efficiency.
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