中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (44): 8237-8242.doi: 10.3969/j.issn.1673-8225.2011.44.018

• 细胞与组织移植 cell and tissue transplantation • 上一篇    下一篇

小鼠胚胎胰腺移植治疗糖尿病

山爱景1，杨  军2，陈  曦2，宁  光3，汪正明1   

  1. 上海交通大学医学院附属瑞金医院，1干细胞移植研究中心，2普外科，3内分泌代谢科，上海市内分泌代谢病研究所，上海市  200025
  • 收稿日期:2011-01-24 修回日期:2011-04-20 出版日期:2011-10-29 发布日期:2011-10-29
  • 通讯作者: 汪正明，博士，上海交通大学医学院附属瑞金医院干细胞移植研究中心，上海市 200025
  • 基金资助:

    国家自然科学基金(30570949, 汪正明)，上海市科委干细胞重大课题基金(06dj14001, 汪正明)，973课题计划(2007CB947901, 汪正明)。

Embryonic mouse pancreas transplantation for treatment of diabetes mellitus

Shan Ai-jing1, Yang Jun2, Chen Xi2, Ning Guang3, Wang Zheng-ming1   

  1. 1Stem Cells Transplantation Research Center, 2Department of Surgery, 3Department of Endocrinology, Shanghai Institute of Endocrinology, Ruijin Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai   200025, China
  • Received:2011-01-24 Revised:2011-04-20 Online:2011-10-29 Published:2011-10-29
  • Contact: Wang Zheng-ming, Ph. D., Stem Cells Transplantation Research Center, Ruijin Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai 200025, China dzmwang@ yahoo.com
  • Supported by:

    following grants to Wand Zheng Ming: the National Natural Science Foundation of China, No. 30570949*; the Major Program of Stem Cells of Shanghai Committee of Science and Technology, No. 06dj14001*; 973 Program of China, No. 2007CB947901*

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摘要：

背景：胚胎胰腺组织具有来源广泛，细胞增殖分化能力强，低免疫排斥性等优点。
目的：探索小鼠胚胎胰腺组织的分离技术，观察其对糖尿病模型小鼠的血糖调节作用。
方法：体视显微镜下分离E11.5~E16.5 C57BL/6小鼠胰腺组织。链唑霉素诱导雄性C57BL/6小鼠建立糖尿病模型，随机分为2组：移植组模型小鼠肾被膜下移植5个E16.5胰腺组织，假手术组模型小鼠肾被膜下注入0.05 mL RPMI1640培养液。移植组小鼠血糖水平≤11.2 mmol/L后，利用IPGTT和IPITT方法检测移植后胚胎胰腺组织的内分泌功能，并摘除移植物观察血糖变化。
结果与结论：体视显微镜下可分离出较完整的E11.5~E16.5小鼠胰腺组织，≤ E12.5 d小鼠胚胎胰腺组织的形态和颜色均难以与周围组织分辨，需根据其与毗邻脏器的关系仔细辨别；> E12.5 d的小鼠胚胎胰腺已初具形态，颜色略发白。组织学和ELISA分别显示胚胎胰腺组织可表达并分泌胰岛素，其表达强度随发育时间逐渐增加。E16.5小鼠胰腺组织移植能有效地控制受体的血糖水平，使受体的体质量和糖耐量恢复正常；胚胎胰腺在受体的肾被膜下可生长发育，摘除的移植物胰岛素和胰高血糖素的表达均较移植前增强。说明胚胎胰腺组织可能成为治疗糖尿病的种子来源。

关键词: 胚胎胰腺, 糖尿病, 动物模型, 分离, 移植, 胰腺干细胞

Abstract:

BACKGROUND: Embryonic pancreatic tissue is characterized by its abundance, potent in proliferation & differentiation, and minimal immunological rejection. It is widely considered as potential pancreatic endocrinological stem cells resource for treating diabetes mellitus.
OBJECTIVE: To investigate the embryonic mouse pancreatic tissue isolation technique and observe the recipients’ blood glucose regulatory effects of the grafted embryonic pancreas in an experimental diabetes mellitus mouse model.
METHODS: Pancreatic tissue from C57BL/6 mouse embryos at embryonic days 11.5-16.5 was isolated under the stereomicroscope. C57BL/6 mouse models of streptozocin-induced diabetes mellitus were established and then randomly divided into two groups: transplantation group, in which, five pieces of pancreatic tissue of mice at embryonic 16.5 days were transplanted into mouse renal capsule, and sham-operated control group, in which, 0.05 mL RPMI1640 culture medium was injected into mouse renal capsule. When blood glucose level of the transplantation group mouse was ≤ 11.2 mmol/L, the endocrine function of embryonic pancreatic tissue transplanted was detected by IPGTT and IPITT methods and then the transplanted graft was removed for observing the blood glucose relapse.
RESULTS AND CONCLUSION: Nearly intact pancreatic tissue of mice at embryonic days 11.5-16.5 could be isolated through the use of stereomicroscope. Pancreatic tissue morphology and color of mice ≤ embryonic 12.5 days were difficultly distinguished from adjacent tissue and they could only be isolated carefully according to the relationship with adjacent organs. Pancreatic tissue of mice > embryonic 12.5 days exhibited initial endocrinological tissue morphology mimic white cauliflower. Histological and ELISA examinations showed that embryonic pancreatic tissue could express and secrete insulin and the insulin level was gradually increased with developmental time. Embryonic pancreatic tissue could grow beneath the recipient renal capsule. The insulin and glucagon expression in the post-transplantational pancreatic tissue graft was increased compared with prior to transplantation. These results suggest that pancreatic tissue is a potential stem cell resource for treating the diabetes mellitus.

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