骨髓基质细胞诱导分化为胰岛素分泌细胞的初步探讨

发布时间：2018-01-08 04:18

本文关键词：骨髓基质细胞诱导分化为胰岛素分泌细胞的初步探讨　出处：《第一军医大学》2005年硕士论文　论文类型：学位论文

【摘要】：【背景】糖尿病是一种慢性使人衰弱且花费昂贵的疾病,它可导致严重的并发症,包括失明及心脏、肾脏等疾病。目前,全世界大约有2亿人在经受着糖尿病的折磨,而且发病人数到2025年为止,将再增长一倍。糖尿病已经成为患者和社会的一个沉重负担,而且问题日益严重,特别是在发展中国家。通过胰腺或胰岛移植治疗1型糖尿病(IDDM)及部分2型糖尿病(NIDDM)是一个很有吸引力的方法,但供体来源不足及免疫排斥反应的存在大大限制了其临床应用。近年来,随着成体干细胞及多能干细胞等概念的提出,通过干细胞移植治疗糖尿病在世界范围内成为一个新的研究热点。骨髓基质细胞(MSCs)是一种存在于骨髓,具有多向分化潜能的非造血细胞,易于分离培养扩增,遗传背景稳定,由于其取材方便、体外扩增能力强、基因转染效率高、转染后稳定表达、可自体回输从而避免了免疫排斥反应,被认为是细胞工程和基因工程理想的种子细胞,而且能克服胚胎干细胞(ESC)所不能解决的伦理学问题。研究表明,在一定条件下,MSCs能分化形成多种组织。【目的】本实验通过体外培养MSCs和胰腺基质细胞,在二者传代稳定后,以胰腺基质细胞培养液诱导培养MSCs,探讨一种在体外条件下将MSCs诱导成为胰岛素分泌细胞(IPCs)的简便方法。【方法】1.取健康成年SD大鼠断髓处死,75%酒精浸泡5min,无菌条件下取出胫骨、股骨,剪去骨骺端,用D-Hanks液冲出骨髓;密度梯度离心法获得单个核细胞,以DMEM/F12全培养基(含碳酸氢钠2.438g/L,胎牛血清100ml/L,青
[Abstract]:[background] Diabetes mellitus is a chronic debilitating and expensive disease that can lead to serious complications, including blindness and heart, kidney and other diseases. About 200 million people worldwide are suffering from diabetes, and the number of cases will double by 2025. Diabetes has become a heavy burden for patients and society. And the problem is increasingly serious, especially in developing countries. The treatment of type 1 diabetes mellitus (IDDM) and partial type 2 diabetes mellitus (NIDDM) through pancreas or islet transplantation is an attractive approach. However, the shortage of donor sources and the existence of immune rejection have greatly limited its clinical application. In recent years, with the development of the concepts of adult stem cells and pluripotent stem cells. Stem cell transplantation has become a new research hotspot in the world. Bone marrow stromal cells (MSCs) are non-hematopoietic cells which exist in bone marrow and have multi-differentiation potential. It is easy to isolate, culture and amplification, stable genetic background, because of its convenient material, in vitro amplification ability, high efficiency of gene transfection, stable expression after transfection, autologous transfusion to avoid immune rejection. It is considered to be an ideal seed cell for cell engineering and genetic engineering, and can overcome the ethical problems that embryonic stem cells cannot solve. MSCs can differentiate into a variety of tissues. [objective] in this experiment, MSCs and pancreatic stromal cells were cultured in vitro and MSCs was induced by pancreatic stromal cell culture medium after stable passage. To explore a simple method for inducing MSCs into insulin-secreting cells in vitro. [methods] 1. Healthy adult SD rats were sacrificed with 75% alcohol for 5 min. Tibia, femur, epiphysis were removed, and bone marrow was washed out with D-Hanks solution. Mononuclear cells were obtained by density gradient centrifugation in DMEM/F12 culture medium (2.438g / L containing sodium bicarbonate, 100ml / L fetal bovine serum, green.
【学位授予单位】：第一军医大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R329

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