چکیده انگلیسی مقاله |
Objective(s): Various microRNAs (miRNAs) are expressed during development of mammalian cells, when they aid in modulating gene expression by mediating mRNA transcript cleavage and/or regulation of translation rate. miR-191 and miR-451 have been shown to be critical regulators of hematopoiesis and have important roles in the induction of erythroid fate decision. So, the aim of this study is investigation of the miR-191 and miR-451 roles in the controlling mouse embryonic stem cell (mESC) differentiation toward the erythroid lineage.Materials and Methods: mESCs were infected with either pCDH-miR-Off-191 viruses in pCDH-miR-Off-191 group or simultaneously with pCDH-miR-Off-191 and pCDH-miR-451 lentiviruses in simultaneous group. Then, the expression profiles of erythroid specific transcription factors and globin genes were analyzed using QRT-PCR on day 14 and 21 of differentiation. Flow cytometry analysis was used to evaluate of TER119 and CD235a erythroid specific surface markers. Results: Gata-1, Klf-1, Epor and globin chains were found to be expressed in pCDH-miR-Off-191 and in simultaneous groups. The majority of globin chains showed changes in their expression levels with progression of differentiation from day 14 to day 21. Flow cytometry results showed that miR-451 up- regulation and miR-191 down-regulation is associated with the expression of TER119 and CD235a. Of these two groups analyzed, simultaneous group was most significantly potent in stimulation of erythroid fate decision of mESCs. Conclusion: Together, present data demonstrate that down-regulation of miR-191 alone can enhance the differentiation of mESCs. However, the simultaneous effect of miR-451up-regulation and miR-191 down-regulation is much stronger and can have more practical use in artificial blood production. |
نویسندگان مقاله |
| Gelareh Shokri Stem Cell Technology Research Center, Tehran, Iran
| Fatemeh Kouhkan Stem Cell Technology Research Center, Tehran, Iran
| shahrzad nojehdehi Stem Cell Technology Research Center, Tehran, Iran
| Masoud Soleimani Department of Hematology, School of Medicine, Tarbiat Modares University, Tehran, Iran
| Ali Akbar Pourfathlah Department of Immunology, School of Medicine, Tarbiat Modares University, Tehran, Iran
| Mahin Nikoguftar Zarif Blood Transfusion Research Center, High Institute for Education and Research in Transfusion Medicine, Tehran, Iran
| Mona Tamadon Stem Cell Technology Research Center, Tehran, Iran
| Narges Obeidi Department of Hematology, School of Para Medicine, Bushehr University of Medical Sciences, Bushehr, Iran
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