Bana Nikoo, Sanooghi Davood, Soleimani Mansoureh, Hayati Roodbari Nasim, Alavi Moghaddam Sepideh, Joghataei Mohammad Taghi, Sayahpour Forough Azam, Faghihi Faezeh
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Department of Genetics, Faculty of Biological Sciences, Shahid Beheshti University, Tehran, Iran.
Tissue Cell. 2017 Aug;49(4):495-502. doi: 10.1016/j.tice.2017.05.003. Epub 2017 May 11.
Musculodegenerative diseases threaten the life of many patients in the world. Since drug administration is not efficient in regeneration of damaged tissues, stem cell therapy is considered as a good strategy to restore the lost cells. Since the efficiency of myogenic differentiation potential of human Chorion- derived Mesenchymal Stem Cells (C-MSCs) has not been addressed so far; we set out to evaluate myogenic differentiation property of these cells in comparison with Umbilical Cord Blood- derived Mesenchymal Stem Cells (UCB-MSCs) in the presence of 5-azacytidine.
MATERIALS & METHODS: To do that, neonate placenta Umbilical Cord Blood were transferred to the lab. After characterization of the isolated cells using flowcytometry and multilineage differentiation capacity, the obtained Mesenchymal Stem Cells were cultured in DMEM/F12 supplemented with 2% FBS and 10μM of 5-azacytidine to induce myogenic differentiation. Real-time PCR and immunocytochemistry were used to assess the myogenic properties of the cells.
Our data showed that C-MSCs and UCB-MSCs were spindle shape in morphology. They were positive for CD90, CD73 and CD44 antigens, and negative for hematopoietic markers. They also differentiated into osteoblast and adipoblast lineages. Real-time PCR results showed that the cells could express MyoD, desmin and α-MHC at the end of the first week (P<0.05). No significant upregulation was detected in the expression of GATA-4 in both groups. Immunocytochemical staining revealed the expression of Desmin, cTnT and α-MHC.
Results showed that these cells are potent to differentiate into myoblast- like cells. An upregulation in the expression of some myogenic markers (desmin, α- MHC) was observed in C-MSCs in comparison with UCB-MSCs.
肌肉退行性疾病威胁着全球众多患者的生命。由于药物给药在受损组织再生方面效果不佳，干细胞疗法被视为恢复丢失细胞的良好策略。鉴于人绒毛膜来源的间充质干细胞（C-MSCs）的成肌分化潜能效率迄今尚未得到研究；我们着手在5-氮杂胞苷存在的情况下，与脐带血来源的间充质干细胞（UCB-MSCs）相比，评估这些细胞的成肌分化特性。
为此，将新生儿胎盘和脐带血转移至实验室。使用流式细胞术和多谱系分化能力对分离的细胞进行表征后，将获得的间充质干细胞培养于添加2%胎牛血清和10μM 5-氮杂胞苷的DMEM/F12中以诱导成肌分化。采用实时聚合酶链反应（PCR）和免疫细胞化学来评估细胞的成肌特性。
我们的数据显示，C-MSCs和UCB-MSCs在形态上呈纺锤形。它们对CD90、CD73和CD44抗原呈阳性，对造血标志物呈阴性。它们还能分化为成骨细胞和成脂细胞谱系。实时PCR结果显示，细胞在第一周结束时可表达MyoD、结蛋白和α-肌球蛋白重链（α-MHC）（P<0.05）。两组中GATA-4的表达均未检测到明显上调。免疫细胞化学染色显示了结蛋白、心肌肌钙蛋白T（cTnT）和α-MHC的表达。
结果表明，这些细胞有分化为成肌样细胞的潜能。与UCB-MSCs相比，C-MSCs中一些成肌标志物（结蛋白、α-MHC）的表达上调。
