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Isolation and culture of adult Sertoli cells and their effects on the function of co-cultured allogeneic islets in vitro

查看全文 作　　者：TENG Yan XUE Wu-jun DING Xiao-ming FENG Xin-shun XIANG He-li JIANG Ya-zhuo TIAN Pu-[1]xun 高影响力作者 机构地区：[1]Department of Kidney Transplantation, First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, China高影响力机构 出　　处：《Chinese Medical Journal》索引2005年第22期,共6页高影响力期刊 基　　金：This study was supported by grants from the Key Program in Scienceand Technique of the National Ministry of Education (No.104169)and the Major Project in Clinical Speciality Foundation of NationalMinistry of Health (Year 2004-2006) 摘　　要：Background Globally, 180 million people suffer from diabetes mellitus. Islet transplantation is believed to be an almost ideal therapy for insulin-dependent patients. How to maintain the viability and the function of isolated human islets is a challenge in clinical practice. Sertoli cells are considered ‘nurse cells’in the seminiferous tubules and have been used in cell graft protocols for neurodegenerative diseases and diabetes in many studies. Many researchers have used immature murine testes as the primarily source of Sertoli cells in islet transplantation because they are easily purified. Mature human Sertoli cells have been seldom investigated. In the present study, we developed a method for the isolation and culture of Sertoli cells derived from adult human testes, and investigated their effects on the function of allogeneic islets when they were cultured together in vitro. Methods Adult Sertoli cells were prepared successfully by two-step enzyme digestion with trypsin, collagenase and hyaluronidase. They were identified by morphological characteristics and their activity was determined by MTT colorimetry over a 28-day culture time in vitro. A glucose-stimulated insulin secretion test was performed to detect the effects of Sertoli cells on allogeneic islets’ function when they were co-cultured for 21 days in vitro. Results In cultured cells, mature human Sertoli cells accounted for more than 90% of total cells. The activity of Sertoli cells reached 95% and they remained highly cytoactive for a long time in vitro (P>0.05). Compared with the islets cultured alone, the co-cultured islets with allogeneic Sertoli cells maintained higher sensitivity to glucose stimulation for the duration of the experiment (P<0.01). Conclusions A method of isolation and culture of Sertoli cells from adult testes has been established. Sertoli cells could enhance allogeneic islets’ function when they were co-cultured in vitro. They could be a helper cell in islet transplantation. 关 键 词：成年 异源组织 胰岛移植 治疗方法
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