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miR-146a and miR-150 promote the differentiation of CD133+ cells into T-lymphoid lineage

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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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Abbreviations

HSC:

Hematopoietic stem cell

qRT-PCR:

Quantitative reverse transcriptase polymerase chain reaction

TCR:

T cell receptor

CD:

Cluster differentiation

UTR:

Untranslated region

miR:

MicroRNA

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Acknowledgments

This study was supported by a Grant from Stem Cell Technology Research Center, Tehran, Iran. Also, we particularly thank Dr. Yousof Gheisari for his scientific assistance in manuscript writing and Fatemeh Kohram for language editing.

Conflict of interest

No competing financial interests exist.

Author information

Authors and Affiliations

  1. Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, 1997775555, Tehran, Iran

    Parviz Fallah, Ehsan Arefian, Mahmood Naderi, Seyed Hamid Aghaee-Bakhtiari, Katayoun Ahmadi & Abbas Shafiee

  2. Department of Pathobiology, School of Medicine, Alborz University of Medical Sciences, Karaj, Iran

    Parviz Fallah

  3. Microbial Biotechnology Laboratory, Department of Microbiology, School of Biology, College of Science, University of Tehran, 14155-6455, Tehran, Iran

    Ehsan Arefian

  4. Department of Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

    Mahmood Naderi

  5. Department of Hematology, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran

    Amir Atashi & Masoud Soleimani

  6. Department of Tissue Engineering, School of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran, Iran

    Abbas Shafiee

Authors
  1. Parviz Fallah
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  2. Ehsan Arefian
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Corresponding author

Correspondence to Masoud Soleimani.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary figure 1: Picture of vector: pLEX-jRED used for cloning miR-146a (TIFF 90 kb)

11033_2013_2567_MOESM2_ESM.tif

Supplementary figure 2: Fluorescent microscopic picture of HEK cells 48 h after transfection (×100). A: Light microscopic picture of the HEK cells. B: The HEK cells transfected with pLEX-jRED + miR-146a. Scale bar=100µm (TIFF 2506 kb)

Supplementary material 3 (DOC 27 kb)

11033_2013_2567_MOESM4_ESM.tif

Supplementary figure 3: Verification of CD133+ cells separated by flow cytometry. Anti-CD133 and anti-CD34 positive cells percent in R1= 97.4% of total cells. RN1= Q2+Q4 = CD34+ cells, RN2 = Q1+Q2 = CD133+ cells (TIFF 243 kb)

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Fallah, P., Arefian, E., Naderi, M. et al. miR-146a and miR-150 promote the differentiation of CD133+ cells into T-lymphoid lineage. Mol Biol Rep 40, 4713–4719 (2013). https://doi.org/10.1007/s11033-013-2567-6

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  • DOI: https://doi.org/10.1007/s11033-013-2567-6

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