Static electromagnetic field and recombinant human fibroblasts encoding miR-451 and miR-16 increased cell trans-differentiation to CD71+ and CD235a+ erythroid like progenitor

Introduction: Ex vivo blood production is an urgent need of most countries, and creating production protocols can save the lives of many patients. Despite the recent advances in blood production in ex vivo conditions, its high-scale production is not yet possible, and requires further studies. There...

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Main Authors: Nafiseh Karoubi, Gholamreza Khamisipour, Nahid Babaei, Narges Obeidi, Abbas Doosti
Format: Article
Language:English
Published: Tabriz University of Medical Sciences 2024-01-01
Series:BioImpacts
Subjects:
Online Access:https://bi.tbzmed.ac.ir/PDF/bi-14-27817.pdf
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author Nafiseh Karoubi
Gholamreza Khamisipour
Nahid Babaei
Narges Obeidi
Abbas Doosti
author_facet Nafiseh Karoubi
Gholamreza Khamisipour
Nahid Babaei
Narges Obeidi
Abbas Doosti
author_sort Nafiseh Karoubi
collection DOAJ
description Introduction: Ex vivo blood production is an urgent need of most countries, and creating production protocols can save the lives of many patients. Despite the recent advances in blood production in ex vivo conditions, its high-scale production is not yet possible, and requires further studies. Therefore, by transfecting fibroblast cells with miR-16, and miR-451 genes, as well as applying low frequency electromagnetic fields (ELF-EMF) treatment, we tried to increase the differentiation of these cells into CD71+ and CD235a+ erythroid like progenitors. Methods: After preparation, and cultivation of human dermal transgenic fibroblast cells, they were transfected by Plenti3-hsa-miR451, Plenti3-hsa-miR16 and Plenti3-backbone inserted into E. coli Stbl4 genome. Then, transgenic fibroblast cells were treated with 10mT ELF-EMF every day for 20 minutes for 7 days. Using a flow cytometer, the expressions of CD71, and CD235a were studied in these cells, and the expressions of genes involved in hematopoiesis were studied using the RT-PCR technique. Results: The results indicated an increase in the differentiation of fibroblast cells treated with 10mT ELF-EMF to erythroid like progenitors. Furthermore, the percentage of CD71+ and CD235a+ cells was the highest in irradiated cells encoding miR-16 and miR-451, which indicates their differentiation into erythroid like progenitors. Also, in the transgenic cells treated with ELF-EMF, an increase in the expressions of α-chain, β-chain, γ-chain and GATA1 genes was observed, which indicates the potential of these cells for hematopoiesis. However, there was no significant difference in the expression of CD34 and CD38 genes in these cell lines. Conclusion: Both ELF-EMF and upregulations of miR-16 and miR-451 lead to improved differentiation of fibroblast cells into erythroid like progenitors.
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spelling doaj.art-dfc1317d82554d0b951ffa63f6505fdc2024-02-06T10:05:09ZengTabriz University of Medical SciencesBioImpacts2228-56522228-56602024-01-01141278172781710.34172/bi.2023.27817bi-27817Static electromagnetic field and recombinant human fibroblasts encoding miR-451 and miR-16 increased cell trans-differentiation to CD71+ and CD235a+ erythroid like progenitorNafiseh Karoubi0Gholamreza Khamisipour1Nahid Babaei2Narges Obeidi3Abbas Doosti4Department of Cell Biology and Genetics, Bushehr Branch, Islamic Azad University, Bushehr, IranDepartment of Cell Biology and Genetics, Bushehr Branch, Islamic Azad University, Bushehr, IranDepartment of Cell Biology and Genetics, Bushehr Branch, Islamic Azad University, Bushehr, IranDepartment of Cell Biology and Genetics, Bushehr Branch, Islamic Azad University, Bushehr, IranDepartment of Cell Biology and Genetics, Bushehr Branch, Islamic Azad University, Bushehr, IranIntroduction: Ex vivo blood production is an urgent need of most countries, and creating production protocols can save the lives of many patients. Despite the recent advances in blood production in ex vivo conditions, its high-scale production is not yet possible, and requires further studies. Therefore, by transfecting fibroblast cells with miR-16, and miR-451 genes, as well as applying low frequency electromagnetic fields (ELF-EMF) treatment, we tried to increase the differentiation of these cells into CD71+ and CD235a+ erythroid like progenitors. Methods: After preparation, and cultivation of human dermal transgenic fibroblast cells, they were transfected by Plenti3-hsa-miR451, Plenti3-hsa-miR16 and Plenti3-backbone inserted into E. coli Stbl4 genome. Then, transgenic fibroblast cells were treated with 10mT ELF-EMF every day for 20 minutes for 7 days. Using a flow cytometer, the expressions of CD71, and CD235a were studied in these cells, and the expressions of genes involved in hematopoiesis were studied using the RT-PCR technique. Results: The results indicated an increase in the differentiation of fibroblast cells treated with 10mT ELF-EMF to erythroid like progenitors. Furthermore, the percentage of CD71+ and CD235a+ cells was the highest in irradiated cells encoding miR-16 and miR-451, which indicates their differentiation into erythroid like progenitors. Also, in the transgenic cells treated with ELF-EMF, an increase in the expressions of α-chain, β-chain, γ-chain and GATA1 genes was observed, which indicates the potential of these cells for hematopoiesis. However, there was no significant difference in the expression of CD34 and CD38 genes in these cell lines. Conclusion: Both ELF-EMF and upregulations of miR-16 and miR-451 lead to improved differentiation of fibroblast cells into erythroid like progenitors.https://bi.tbzmed.ac.ir/PDF/bi-14-27817.pdfelf-emferythroidfibroblastgenehematopoiesis
spellingShingle Nafiseh Karoubi
Gholamreza Khamisipour
Nahid Babaei
Narges Obeidi
Abbas Doosti
Static electromagnetic field and recombinant human fibroblasts encoding miR-451 and miR-16 increased cell trans-differentiation to CD71+ and CD235a+ erythroid like progenitor
BioImpacts
elf-emf
erythroid
fibroblast
gene
hematopoiesis
title Static electromagnetic field and recombinant human fibroblasts encoding miR-451 and miR-16 increased cell trans-differentiation to CD71+ and CD235a+ erythroid like progenitor
title_full Static electromagnetic field and recombinant human fibroblasts encoding miR-451 and miR-16 increased cell trans-differentiation to CD71+ and CD235a+ erythroid like progenitor
title_fullStr Static electromagnetic field and recombinant human fibroblasts encoding miR-451 and miR-16 increased cell trans-differentiation to CD71+ and CD235a+ erythroid like progenitor
title_full_unstemmed Static electromagnetic field and recombinant human fibroblasts encoding miR-451 and miR-16 increased cell trans-differentiation to CD71+ and CD235a+ erythroid like progenitor
title_short Static electromagnetic field and recombinant human fibroblasts encoding miR-451 and miR-16 increased cell trans-differentiation to CD71+ and CD235a+ erythroid like progenitor
title_sort static electromagnetic field and recombinant human fibroblasts encoding mir 451 and mir 16 increased cell trans differentiation to cd71 and cd235a erythroid like progenitor
topic elf-emf
erythroid
fibroblast
gene
hematopoiesis
url https://bi.tbzmed.ac.ir/PDF/bi-14-27817.pdf
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