摘要
目的:前期实验采用药物诱导的方法将人脐带间充质干细胞诱导分化为胰岛样细胞,将人脐带间充质干细胞与大鼠胰岛细胞共同培养及移植入糖尿病大鼠体内,观察其在胰岛细胞生长的微环境及在体环境中向胰岛样细胞分化的潜能。方法:实验于2006-10/2007-09在汕头大学医学院生殖遗传实验室完成,属于广东省科技厅重点实验室。①实验材料:SD大鼠由广州中医药大学实验动物中心提供。对动物处置符合动物伦理学标准。脐带取自汕大附二妇产科健康足月妊娠剖宫产的胎儿,产妇及家属对实验知情同意,并经医院伦理委员会批准。②实验方法:分离培养人脐带间充质干细胞,并采用酶消法分离大鼠胰岛细胞。将人脐带间充质干细胞与大鼠胰岛细胞以半透膜相隔共同培养,在共培养的第3,7,14天采用放射免疫法检测胰岛素水平,RT-PCR方法检测PDX-1基因的表达,以单独培养的脐带间充质干细胞为对照。经尾静脉将人脐带间充质干细胞移植入糖尿病模型大鼠体内,采用半定量RT-PCR方法检测人insulin基因表达。结果:①共培养条件下的人脐带间充质干细胞由长梭形逐渐变圆,并聚集成团。②放射免疫法结果表明,共培养组人脐带间充质干细胞随葡萄糖浓度的增高而分泌的胰岛素量也增加,与对照组相比差异显著(P<0.05)。③未经共培养诱导的人脐带间充质干细胞不表达PDX-1,共培养3d的人脐带间充质干细胞表达PDX-1,共培养7d的人脐带间充质干细胞仍表达PDX-1,共培养14d的人脐带间充质干细胞未表达PDX-1。④未经移植人脐带间充质干细胞的大鼠胰腺不表达人insulin基因,而移植人脐带间充质干细胞的大鼠胰腺在第60天时能够检测出人insulin基因。结论:人脐带间充质干细胞具有在体内外的微环境中向胰岛样细胞分化的潜能。
AIM： Human umbilical cord mesenchymal stem cells （MSCs） were induced into islet-like cells by medicine during our prophase research. We cocutured MSCs with pancreatic cells of neonatal rat and transplanted MSCs into the body of diabetic rat. This study explored the possibility of differentiation of inducing human umbilical cord MSCs into islet-like cells in microenvironment of pancreatic cells of neonatal rat and in body of diabetic rat. METHODS： Experiments were performed from October 2006 to September 2007 at the Reproduction Genetic Laboratory of Medical College of Shantou University. SD rats were bought from Experimental Animal Center of Guangzhou University of Chinese Medicine. According to the animal ethical standards, the animals were disposed in the experiment. The human umbilical cords were obtained from healthy caesarean delivered uterogestation foetus from the Second Affdiated Hospital of Shantou University. The informed consents were obtained from the lying-in mother and her family. The experiment was approved by Hospital＇s Ethics Committee. The pancreatic cells were isolated from fetal rats by enzyme dispelling therapy and human umbilical cord MSCs were also isolated and cultured, and then cocultured with rat pancreatic cells. On the third, 7^th and 14^th days during coculture, insulin levels were detected by radio-immunity. Human pancreatic and duodenal homeobox I（PDX-I） was detected with reverse transcription-polymerase chain reaction （RT-PCR） method. Alone cultured MSCs was considered to be a control group. Human umbilical cord MSCs were transplanted into the body of diabetic rats through vena caudalis, then human insulin gene was detected by RT-PCR.
RESULTS： The morphology of human umbilical cord MSCs under coculturing gradually changed from fibroblast to round and had the tendency of forming clusters. The results of radio-immunity manifested that the insulin levels secreted by cocultured MSCs could increase following an increase in glucose concentration. There were signific
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第21期4102-4106,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
广东省科技计划项目(2006B3600502)“人脐带间充质干细胞对Ⅰ型糖尿病治疗作用的研究”
广东省科技计划项目(2007B031503007)“双基因修饰huMSCs的MRI在体显像示踪和治疗Ⅰ型糖尿病的研究”~~
