Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
Department of Hematology and Medical Laboratory Sciences, Faculty of Allied Medicine, Kerman University of Medical Sciences, Kerman, Iran.
Biochem Genet. 2024 Aug;62(4):2796-2809. doi: 10.1007/s10528-023-10565-9. Epub 2023 Nov 29.
Several investigations are being done to increase the short lifetime of mesenchymal stem cells (MSCs). One of the crucial genes involved in the immortalization of MSCs, hTERT (human telomerase reverse transcriptase), is activated in most publications using viral-based techniques. In this work, we investigated the use of platelet-derived (PMPs) and B cell precursor leukemia-derived microparticles as a nonviral method to trigger and compare the expression of the hTERT gene in MSCs. MSCs were extracted from the umbilical cord for the current investigation and identified using a flow cytometry approach and an inverted microscope. The Nalm-6 cell line and platelet concentrate were used to isolate microparticles (MPs). MSCs and MPs were cocultured for 14 days at 25-, 50-, and 100 μg/ml concentrations. qRT-PCR was used to research the expression of the hTERT gene. SPSS 26.0's t test was used to compare the outcomes. After coculture with platelet MPs, MSCs had higher levels of hTERT gene expression than the control group. In contrast, this gene's expression was concurrently decreased in MSCs exposed to MPs generated from Nalm-6. We demonstrated that following 14-day treatment, PMP significantly boosted the hTERT gene expression in MSCs, while the Nalm-6 MPs lowered the gene expression. However, additional studies are necessary due to the stability of hTERT gene expression and the immortalization of MSCs following exposure.
目前正在进行多项研究以延长间充质干细胞(MSCs)的短期寿命。在大多数使用基于病毒的技术进行的研究中,涉及 MSC 永生化的关键基因之一 hTERT(人端粒酶逆转录酶)被激活。在这项工作中,我们研究了使用血小板衍生(PMPs)和 B 细胞前体细胞白血病衍生的微颗粒作为非病毒方法来触发和比较 MSC 中 hTERT 基因的表达。为了进行当前的研究,从脐带中提取了 MSC,并使用流式细胞术和倒置显微镜进行了鉴定。使用 Nalm-6 细胞系和血小板浓缩物来分离微颗粒(MPs)。将 MSC 和 MPs 在 25、50 和 100μg/ml 浓度下共培养 14 天。使用 qRT-PCR 研究 hTERT 基因的表达。使用 SPSS 26.0 的 t 检验来比较结果。与对照组相比,与血小板 MPs 共培养后,MSC 中 hTERT 基因的表达水平更高。相比之下,暴露于源自 Nalm-6 的 MPs 的 MSC 中该基因的表达同时降低。我们证明,经过 14 天的治疗后,PMP 可显著增强 MSC 中 hTERT 基因的表达,而 Nalm-6 MPs 则降低了该基因的表达。然而,由于 hTERT 基因表达的稳定性以及暴露后 MSC 的永生化,还需要进一步的研究。