Ebnerasuly Farid, Hajebrahimi Zahra, Tabaie Seyed Mehdi, Darbouy Mojtaba
Department of Biology, Fars Science and Research Branch , Islamic Azad University, Marvdasht, Iran.
Department of Biology, Marvdasht Branch, Islamic Azad University, Marvdasht, Iran.
Iran J Biotechnol. 2017 Dec 29;15(4):241-251. doi: 10.15171/ijb.1747. eCollection 2017.
Mesenchymal stem cells (MSCs) are multipotent cells able to differentiating into a variety of mesenchymal tissues including osteoblasts, adipocytes and several other tissues. Differentiation of MSCs into fibroblast cells is an attractive strategy to achieve fibroblast cell and use them for purposes such as regeneration medicine. The goal of this study was investigate the simulated microgravity effect on differentiation of Adipose Derived Stem Cells (ADSCs) to fibroblasts. To fibroblast differentiation 100 ng.mL of connective tissue growth factor (CTGF), and for simulation microgravity, 2D clinostat was used. After isolation the human ADSCs from adipose, cells were passaged, and at passages 3 they were used for characterization and subsequent steps. After 7 days of CTGF and simulated microgravity treatment, proliferation, and differentiation were analyzed collectively by MTT assay, quantitative PCR analyses, and Immunocytochemistry staining. MTT assay revealed that CTGF stimulate the proliferation but simulated microgravity didn't have statistically significant effect on cell proliferation. In RNA level the expression of these genes are investigated: collagen type I (), elastin (), collagen type III (), Matrix Metalloproteinases I(), Fibronectin 1 (), CD44, Fibroblast Specific protein (), Integrin Subunit Beta 1 (), Vimentin () and Fibrillin (). We found that expression of , FN1, , , , and in both condition, and VIM and FBN1 just in differentiation medium in normal gravity increased. In protein level the expression of COL III and ELN in simulated microgravity increased. These findings collectively demonstrate that the simulated microgravity condition alters the marker fibroblast gene expression in fibroblast differentiation process.
间充质干细胞（MSCs）是多能细胞，能够分化为多种间充质组织，包括成骨细胞、脂肪细胞和其他几种组织。将MSCs分化为成纤维细胞是一种有吸引力的策略，可获得成纤维细胞并将其用于再生医学等目的。本研究的目的是研究模拟微重力对脂肪来源干细胞（ADSCs）向成纤维细胞分化的影响。为了诱导成纤维细胞分化，使用了100 ng/mL的结缔组织生长因子（CTGF），为了模拟微重力，使用了二维回转器。从脂肪中分离出人ADSCs后，细胞进行传代培养，在第3代时用于表征和后续步骤。经过7天的CTGF和模拟微重力处理后，通过MTT法、定量PCR分析和免疫细胞化学染色共同分析细胞增殖和分化情况。MTT分析表明，CTGF刺激细胞增殖，但模拟微重力对细胞增殖没有统计学上的显著影响。在RNA水平上，研究了这些基因的表达：I型胶原蛋白（）、弹性蛋白（）、III型胶原蛋白（）、基质金属蛋白酶I（）、纤连蛋白1（）、CD44、成纤维细胞特异性蛋白（）、整合素亚基β1（）、波形蛋白（）和原纤蛋白（）。我们发现，在两种条件下，、FN1、、、和的表达均有变化，而VIM和FBN1仅在正常重力下的分化培养基中表达增加。在蛋白质水平上，模拟微重力下COL III和ELN的表达增加。这些发现共同表明，模拟微重力条件在成纤维细胞分化过程中改变了成纤维细胞标记基因的表达。
