袁丽丽,张炳强,王玮玮,李蒙,张嘉澍,胡鑫童.人脐带间充质干细胞体外诱导分化胰岛样细胞及形态学观察[J].济宁医学院学报,2017,40(2):93-97,102 |
人脐带间充质干细胞体外诱导分化胰岛样细胞及形态学观察 |
Morphological study of umbilical cord derived mesenchymal stem cells differentiating into islet-like cells in vitro |
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DOI:10.3969/j.issn.1000-9760.2017.02.004 |
中文关键词: 人脐带间充质干细胞;分化;胰岛样细胞 |
英文关键词: HUMSCs;Differentiation;Islet-like cell |
基金项目:济宁医学院教师指导学生科研项目(2015ZDXS-8) |
作者 | 单位 | 袁丽丽 | 济宁医学院基础医学院, 济宁 272067 | 张炳强 | 卫生部细胞移植重点实验室临床中心, 青岛 266000 | 王玮玮 | 济宁医学院, 济宁 272029 | 李蒙 | 济宁医学院, 济宁 272029 | 张嘉澍 | 济宁医学院, 济宁 272029 | 胡鑫童 | 济宁医学院, 济宁 272029 |
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中文摘要: |
目的 构建人脐带间充质干细胞(human umbilical cord derived mesenchymal stem cells,HUMSCs)体外培养体系,并诱导其向胰岛样细胞分化。方法 体外分离培养HUMSCs,采用流式细胞术(flow cytometry,FCM)对HUMSCs特异标记物进行检测;先后加入β-巯基乙醇及高糖的分步诱导法诱导HUMSCs向胰岛样细胞分化,采用RT-PCR、双硫腙染色对诱导后细胞进行胰岛样细胞鉴定。结果 FCM结果显示HUMSCs表达CD29、CD73、CD90及CD105,不表达或低表达CD45、CD86、HLA-DR及CD34;诱导的胰岛样细胞表达PDX-1和Insulin,双硫腙染色阳性。结论 HUMSCs可经分步诱导法分化为胰岛样细胞,能为细胞移植治疗糖尿病提供丰富的种子细胞。 |
英文摘要: |
Objective To establish a cell line of human umbilical cord derived mesenchymal stem cells (HUMSCs) and induce them differentiating into islet-like cells in vitro.Methods In vitro,mesenchymal stem cells were isolated and cultivated from human umbilical cord.Flow cytometry (FCM) was used to analyze the specific markers of mesenchymal stem cells.The HUMSCs were induced to differentiating into islet-like cells conducted by stepwise,firstly β-mercaptoethanol,and then high glucose.RT-PCR and dithizone staining were applied for identifying the islet-like cells.Results CD29,CD73,CD90,and CD105 were expressed in the mesenchymal stem cells,while there was no expression or low expression of CD45,CD86,HLA-DR,CD34.PDX-1 and Insulin were detected by RT-PCR.The dithizone staining was positive in the induced cells.Conclusion It shows a stable cell line of HUMSCs,which successfully gains islet-like cells by the following induced treatments of β-mercaptoethanol and high glucose.It could provide abundant seeding cells for cell transplantation on treating diabetes. |
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