上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2013, Vol. 33 ›› Issue (6): 763-.doi: 10.3969/j.issn.1674-8115.2013.06.013

• 论著（基础研究） • 上一篇    下一篇

新鲜与短期培养胰岛细胞移植对移植胰岛存活的影响

唐成佳，杜成友，刘定志，潘 龙   

  1. 重庆医科大学附属第一医院肝胆外科， 重庆 400016
  • 出版日期:2013-06-28 发布日期:2013-06-28
  • 通讯作者: 杜成友, 电子信箱: duchengyou@126.com。
  • 作者简介:唐成佳（1985—）， 男， 硕士生; 电子信箱: lxwxljtang@163.com。
  • 基金资助:

    国家自然科学基金（30972945）和重庆市科委基金（CSTC2007 BB5263）

Influence of fresh and short-term cultured islet transplantation on survival of transplanted islets

TANG Cheng-jia, DU Cheng-you, LIU Ding-zhi, PAN Long   

  1. Department of Hepatobiliary Surgery, the First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China
  • Online:2013-06-28 Published:2013-06-28
  • Supported by:

    National Natural Science Foundation of China, 30972945; Chongqing Science and Technology Committee Foundation, CSTC2007 BB5263

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摘要：

目的 观察新鲜和短期培养后的小鼠胰岛细胞对移植后胰岛的再血管化及功能的影响。方法 C57小鼠作为胰腺供体，胆总管灌注胶原酶P溶液消化胰腺，Ficoll 400不连续梯度纯化获得胰岛细胞。接受不同胰岛细胞肾被膜下移植的受体糖尿病C57小鼠随机分成三组（n=10）：A组接受新鲜胰岛细胞移植，B、C组分别接受培养1、3 d的胰岛细胞移植。术后观察血糖变化并于移植后第14天取移植胰岛，分别进行HE染色和胰岛素、CD31免疫组织化学染色，并计算微血管密度。结果 胰岛细胞移植后3 d内，三组小鼠的随机血糖浓度均<11.0 mmol/L。A组小鼠在移植后第14天时的血糖浓度仍<11.0 mmol/L，而B、C组小鼠血糖浓度在移植3 d后呈现持续上升，此后各时间点的血糖浓度均显著高于A组，差异有统计学意义（P<0.05）。免疫组织化学胰岛素染色结果显示：A组小鼠肾被膜下成团的细胞被染成棕褐色，B、C组类似细胞团仅有少量着色。免疫组织化学CD31染色发现，A组小鼠肾被膜下移植细胞团内部有大量细胞胞质被染成棕黄色，而B组和C组仅有少量细胞着色；A组肾被膜下胰岛内微血管密度显著高于B、C组（P<0.05）。结论 小鼠新鲜胰岛细胞移植较短期培养后胰岛细胞移植更有利于移植胰岛的再血管化及存活。

关键词: 胰岛移植, 胰岛细胞培养, 新鲜移植, 糖尿病小鼠

Abstract:

Objective To investigate the influence of fresh and short-term cultured islets on revascularization and function of transplanted islets in mice. Methods Pancreatic islets of donors were obtained from C57 mice by digestion of collagenase P solution and Ficoll 400 density gradient centrifugation. The diabetic C57 mice undergoing syngenetic islet transplantation under renal capsule were randomly divided into three groups, with 10 mice in each group. Mice in group A were transplanted with fresh islets, and those in group B and group C were transplanted with islets with culture of 1 d and 3 d respectively. The changes of blood glucose in diabetic mice after transplantation were observed. The transplanted islets were obtained 14 d after transplantation, HE staining was conducted, immunohistochemical staining of insulin and CD31 was performed, and the microvascular density was calculated. Results The blood glucose of mice was lower than 11.0 mmol/L within 3 d after islet transplantation in three groups. The blood glucose of mice in group A maintained less than 11.0 mmol/L on the 14th day after transplantation, while the blood glucose in group B and group C continued to rise 3 d after transplantation, and was significantly higher than that in group A since then (P<0.05). Immunohistochemical staining of insulin demonstrated that there were a large number of insulin positive cells in cell masses in group A, whereas insulin positive cells were seldomly found in group B and group C. Immunohistochemical staining of CD31 demonstrated a large number of cells positively stained by CD31 under the renal capsule in group A, while there were fewer positive cells in group B and group C. The microvascular density in islets in group A was significantly higher than those in group B and group C (P<0.05). Conclusion Fresh islet transplantation is better than short-term cultured islet transplantation in improving the revascularization and survival of transplanted islets in mice.

Key words: islet transplantation, islet culture, fresh infusion, diabetic mice