小鼠胰腺导管上皮细胞和骨髓间充质干细胞转分化为胰岛样细胞的实验研究

发布时间：2018-04-02 14:36

本文选题：胰岛　切入点：导管细胞　出处：《重庆医科大学》2005年博士论文

【摘要】：目的: 1 型糖尿病的发生是由于胰岛素的绝对缺乏,需终身依赖胰岛素治疗,但是胰岛素治疗并不能完全阻止糖尿病并发症的发生,因此寻找合适的细胞以替代β细胞发挥生理功能具有重要的意义。Shapiro 等报道,给 7 例 1 型糖尿病患者移植新鲜胰岛并配合非糖皮质激素免疫抑制剂治疗,患者不依赖外源胰岛素而血糖控制良好的平均时间超过了 12 个月,可是,供体来源的匮乏以及移植免疫排斥反应极大地阻碍了该方法的推广和应用。体外培养干细胞使其定向分化为胰岛样细胞以替代患者体内受损胰岛的生理功能,可能是解决供体来源匮乏以及移植免疫排斥反应有效方法之一。研究证实小鼠胰腺导管上皮细胞能够分化为具有内分泌功能的胰岛样细胞, 本研究采用细胞培养及诱导分化技术,分离培养小鼠胰腺导管上皮细胞并使之转分化为胰腺干细胞及胰岛素分泌细胞(胰岛样细胞),并将胰岛样细胞移植入 1 型糖尿病小鼠体内,定期观察血糖水平 2 个月,为胰岛细胞移植治疗糖尿病奠定实践基础。材料与方法: 1、小鼠胰腺导管上皮细胞原代培养、诱导分化及鉴定:实验用成年昆明小鼠,以Ⅴ型胶原酶消化加滤网过滤法,分离小鼠胰腺导管上皮细胞,用添加有多种生长因子的 DMEM/F12 培养基培养并诱导分
[Abstract]:Objective:Type 1 diabetes occurs because of an absolute lack of insulin, which requires life-long insulin therapy, but insulin therapy does not completely prevent the occurrence of diabetic complications.Therefore, it is important to find suitable cells to substitute 尾 cells for physiological function. Shapiro et al reported that 7 patients with type 1 diabetes were transplanted with fresh pancreatic islets and treated with non-glucocorticoid immunosuppressant.The average time for patients to control their blood sugar well without exogenous insulin is more than 12 months. However, the lack of donor sources and allograft immune rejection have greatly hindered the development and application of this method.In vitro culture of stem cells to differentiate them into islet like cells to replace the physiological function of damaged islets in patients may be one of the effective methods to solve the shortage of donor sources and the transplantation of immune rejection.It was proved that the pancreatic ductal epithelial cells of mice could differentiate into islet like cells with endocrine function. In this study, the technique of cell culture and differentiation was used to induce the differentiation of pancreatic ductal epithelial cells.Pancreatic ductal epithelial cells were isolated and cultured and transformed into pancreatic stem cells and insulin secreting cells (islet like cells), and islet like cells were transplanted into type 1 diabetic mice. Blood glucose levels were observed regularly for 2 months.To lay a practical foundation for the treatment of diabetes mellitus by islet cell transplantation.Materials and methods:1. Primary culture, differentiation and identification of pancreatic ductal epithelial cells in mice. The pancreatic ductal epithelial cells were isolated by collagenase V digestion and filter filtration in adult Kunming mice.Culture and Induction of DMEM/F12 medium with multiple growth factors
【学位授予单位】：重庆医科大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R329.2

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