Rahimi Maryam, Zarnani Amir-Hassan, Mohseni-Kouchesfehani Homa, Soltanghoraei Haleh, Akhondi Mohammad-Mehdi, Kazemnejad Somaieh
Department of Animal Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.
Mol Biotechnol. 2014 Dec;56(12):1151-62. doi: 10.1007/s12033-014-9795-4.
In recent years, menstrual blood-derived stem cells (MenSCs) have been introduced as easily accessible and refreshing stem cell source without ethical considerations in the field of regenerative medicine. The aim of this study was to investigate in vitro cardiac differentiation capacity of MenSCs compared to bone marrow-derived stem cells (BMSCs) under two protocols using 5-aza-2'-deoxycytidine (5-aza) and basic fibroblast growth factor (bFGF). Our data revealed that differentiated MenSCs and BMSCs acquired some features of cardiomyocytes; however, degree of differentiation was dependent on the protocol. In a similar manner with BMSCs, differentiated MenSCs showed upper levels of mRNA/protein of late-stage cardiac markers under 5-aza stimulation and continuous treatment with bFGF (protocol 2) compared to those induced by 5-aza alone (protocol 1) evidencing the key role of bFGF in cardiac development of stem cells. Compared to corresponding undifferentiated cells differentiated MenSCs under protocol 2 showed remarkable expression of connexin-43 and TNNT2 at both gene and protein levels, whereas developed BMSCs under the same condition only expressed connextin-43 at the higher level. Superiority of protocol 2 over protocol 1 was confirmed by assessment of LDH and cTnI production by differentiated cells. Based on the accumulative data, our study provided convincing evidence that MenSCs have relatively higher capability to be differentiated toward cardiomyocyte compared with BMSCs. Furthermore, usage of bFGF and 5-aza to induce in vitro cardiac differentiation of MenSCs is highly recommended.
近年来,月经血源性干细胞(MenSCs)作为一种易于获取且不受伦理限制的干细胞来源,被引入到再生医学领域。本研究的目的是在两种使用5-氮杂-2'-脱氧胞苷(5-aza)和碱性成纤维细胞生长因子(bFGF)的方案下,比较MenSCs与骨髓源性干细胞(BMSCs)的体外心肌分化能力。我们的数据显示,分化后的MenSCs和BMSCs获得了一些心肌细胞的特征;然而,分化程度取决于方案。与BMSCs类似,与单独使用5-aza诱导(方案1)相比,在5-aza刺激和持续使用bFGF处理(方案2)下,分化后的MenSCs在晚期心脏标志物的mRNA/蛋白水平上更高,这证明了bFGF在干细胞心脏发育中的关键作用。与相应的未分化细胞相比,方案2下分化后的MenSCs在基因和蛋白水平上均显著表达连接蛋白-43(connexin-43)和肌钙蛋白T2(TNNT2),而在相同条件下分化后的BMSCs仅在较高水平表达连接蛋白-43。通过评估分化细胞的乳酸脱氢酶(LDH)和心肌肌钙蛋白I(cTnI)产量,证实了方案2优于方案1。基于累积数据,我们的研究提供了令人信服的证据,表明与BMSCs相比,MenSCs向心肌细胞分化的能力相对更高。此外,强烈推荐使用bFGF和5-aza诱导MenSCs的体外心肌分化。