Jalili Angourani Khadijeh, Mazhari Sogol, Farivar Shirin, Salman Mahini Donya, Rouintan Abdolreza, Baghaei Kaveh
Department of Molecular and Cell Biology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran.
Basic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Science, Tehran, Iran.
Gastroenterol Hepatol Bed Bench. 2018 Winter;11(Suppl 1):S73-S79.
The aim of the present study was to investigate the effect of human bone marrow-derived mesenchymal stem cells conditioned medium on fibroblast to myofibroblast differentiation.
Mesenchymal stem cells have a long-term clinical application and widely have used in autoimmune disease and regenerative medicine. However, some MSCs derived cytokines such as TGF-β could have a dual role in suppression or progression of disease. Fibroblast activation and extracellular matrix production are two key features of wound healing which mostly are controlled with multifunctional cytokine TGF-β1.
Bone marrow MSCs were isolated, cultured and used for conditioned medium preparation. The flow cytometry analysis was done for MSCs cell surface markers. MRC-5 subconfluent cells were starved with the medium containing 0.5 % FBS for 24h, then treated with exogenous TGF-β1 (10ng/ml as positive control) and MSCs-conditioned medium for 48h. Finally, the mRNA expression of three target genes: collagen I, collagen III and α-SMA were evaluated by RT-PCR technique.
Our findings demonstrated that bone marrow-derived mesenchymal stem cells-conditioned medium (secretome) significantly upregulated type I and III collagen expression but non-significantly α-SMA gene expression.
Totally, Real Time PCR results suggest that MSCs conditioned medium activates differentiation of fibroblast to myofibroblast phenotype as confirmed through the presence of α-SMA, collagen I and collagen III expression compared to control in MRC 5 cells.
本研究旨在探讨人骨髓间充质干细胞条件培养基对成纤维细胞向肌成纤维细胞分化的影响。
间充质干细胞具有长期临床应用价值,广泛应用于自身免疫性疾病和再生医学。然而,一些间充质干细胞衍生的细胞因子,如转化生长因子-β,在疾病的抑制或进展中可能具有双重作用。成纤维细胞活化和细胞外基质产生是伤口愈合的两个关键特征,主要由多功能细胞因子转化生长因子-β1控制。
分离、培养骨髓间充质干细胞并用于制备条件培养基。对间充质干细胞的细胞表面标志物进行流式细胞术分析。将MRC-5亚汇合细胞用含0.5%胎牛血清的培养基饥饿培养24小时,然后用外源性转化生长因子-β1(10纳克/毫升作为阳性对照)和间充质干细胞条件培养基处理48小时。最后,通过逆转录-聚合酶链反应技术评估三个靶基因:I型胶原、III型胶原和α-平滑肌肌动蛋白的mRNA表达。
我们的研究结果表明,骨髓来源的间充质干细胞条件培养基(分泌组)显著上调I型和III型胶原表达,但α-平滑肌肌动蛋白基因表达无显著变化。
总体而言,实时聚合酶链反应结果表明,间充质干细胞条件培养基激活了成纤维细胞向肌成纤维细胞表型的分化,这通过与MRC 5细胞中的对照相比,α-平滑肌肌动蛋白、I型胶原和III型胶原的表达得以证实。