■离体血液生产是大多数国家的迫切需要,创建生产协议可以挽救许多患者的生命。尽管最近在离体条件下的血液生产方面取得了进展,它的大规模生产还不可能,需要进一步研究。因此,通过用miR-16和miR-451基因转染成纤维细胞,以及应用低频电磁场(ELF-EMF)治疗,我们试图增加这些细胞向CD71+和CD235a+红系样祖细胞的分化。
■准备后,和培养人真皮转基因成纤维细胞,它们被Plenti3-hsa-miR451、Plenti3-hsa-miR16和Plenti3-backbone插入大肠杆菌Stbl4基因组中转染。然后,转基因成纤维细胞每天用10mTELF-EMF处理20分钟,持续7天。用流式细胞仪,研究了CD71和CD235a在这些细胞中的表达,并使用RT-PCR技术研究了与造血有关的基因的表达。
■结果表明用10mTELF-EMF处理的成纤维细胞向红系祖细胞的分化增加。此外,在编码miR-16和miR-451的辐射细胞中,CD71+和CD235a+细胞的百分比最高,这表明它们分化为红细胞样祖细胞.此外,在用ELF-EMF处理的转基因细胞中,α链表达的增加,β-链,观察到γ链和GATA1基因,这表明了这些细胞造血的潜力。然而,CD34和CD38基因在这些细胞系中的表达没有显著差异。
■ELF-EMF和miR-16和miR-451的上调均导致成纤维细胞向红系样祖细胞分化的改善。
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: Both ELF-EMF and upregulations of miR-16 and miR-451 lead to improved differentiation of fibroblast cells into
erythroid like progenitors.