The effect of dimethyl sulfoxide on hepatic differentiation of mesenchymal stem cells
The effect of dimethyl sulfoxide on hepatic differentiation of mesenchymal stem cells
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DOI：
10.3109/21691401.2014.928778
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发表时间：
2016-01-02
影响因子：
5.8
通讯作者：
Mohammadi, Seyed Abolghasem
中科院分区：
文献类型：
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作者：
Alizadeh, Effat;Zarghami, Nosratollah;Mohammadi, Seyed Abolghasem
简介：脂肪组织源性间充质干细胞（AT-MSCs）由于其肝分化潜力，是自体干细胞治疗肝脏相关疾病的合适选择。二甲亚砜 (DMSO) 是一种两亲性分子，具有将亲脂性和亲水性试剂输送到细胞中的潜力，也是用于冷冻细胞的常见冷冻保护剂。一些方案中使用 DMSO 将 AT-MSC 诱导为肝细胞样细胞。然而，此前并未调查DMSO对AT-MSCs肝源分化的影响。在本研究中，我们旨在评估DMSO对AT-MSCs向肝谱系分化的影响。方法：我们从脂肪组织中分离间充质干细胞（MSCs），然后验证AT-MSCs的多效性和表面标记。分离的AT-MSCs随机分四组，第1组：HGF处理，第2组：HGF+DMSO处理，第3组：HGF+DMSO+OSM处理，对照组在无血清扩增培养基中培养3周； EGF 和 bFGF 也包含在诱导的第一天。用显微镜观察诱导期的形态变化。使用 ELISA 研究分化 MSC 的白蛋白 (ALB) 分泌，并使用比色测定评估尿素产量。 qRT-PCR 用于定量肝细胞标记基因，包括 AFP、ALB、CK18、HNF4a 和 HNF6。通过高碘酸希夫氏染色观察分化细胞的糖原储存。结果：结果表明，DMSO 加速了 AT-MSC 的肝分化，其特征是形态快速变化；肝脏标志基因（ALB）在mRNA和蛋白水平上均较高表达（P < 0.05）；还增加了其他肝脏基因的转录水平，包括 CK18、HNF4a 和 HNF6（P < 0.01）；此外，DMSO 处理组的糖原储存百分比更高（p < 0.05）。结论：DMSO 催化肝分化；因此，使用 DMSO 加速 AT-MSC 的肝生成方案似乎是有利的。
Introduction: Adipose tissue-derived mesenchymal stem cells (AT-MSCs) are suitable choices in autologous stem cell treatment of liver-associated diseases due to their hepatic differentiation potential. Dimethyl sulfoxide (DMSO) is an amphipathic molecule with potential of delivering both lipophilic and hydrophilic agents into cells, also a common cryoprotectant for freezing of the cells. DMSO was used in some protocols for induction of AT-MSCs towards hepatocyte like cells. However, the effect of DMSO on hepatogenic differentiation of AT-MSCs were not surveyed, previously. In the present study, we aimed at evaluation of the effect of DMSO on differentiation of AT-MSCs into hepatic lineage.Methods: We isolated mesenchymal stem cells (MSCs) from adipose tissue, and then verifies multi-potency and surface markers of AT-MSCs. Isolated AT-MSCs randomly dispensed in four groups including Group 1: HGF treated, 2: HGF + DMSO treated, 3: HGF + DMSO + OSM treated, and group control for a period of 3 weeks in the expansion medium without serum; EGF and bFGF were also included in the first days of inductions. The morphologic changes during induction period was observed with microscopy. The secretion of albumin (ALB) of the differentiating MSCs was investigated using ELISA, and urea production was evaluated using colorimetric assay. The qRT-PCR was performed for quantitation of hepatocyte marker genes including AFP, ALB, CK18, HNF4a, and HNF6. The glycogen storage of differentiated cells was visualized by periodic-acid Schiff's staining.Results: The results demonstrate that DMSO speeds up hepatic differentiation of AT-MSCs characterized by rapid changes in morphology; higher expression of hepatic marker gene (ALB) in both mRNA and protein level (P < 0.05); also increased transcriptional levels of other liver genes including CK18, HNF4a, and HNF6 (P < 0.01); and moreover, greater percentage of glycogen storage(p < 0.05) in DMSO-treated groups.Conclusion: DMSO catalyzes hepatic differentiation; therefore, using DMSO for acceleration of the hepatogenic protocols of AT-MSCs appears advantageous.