GFP标记胰岛及其在鼠体内不同部位移植的探索  

Exploration of GFP labeled pancreatic islets and their transplantation in different sites of mice and rats

作　　者：蔡娟娟 叶泽岚 宋益康 林悦 傅红兴 CAI Juanjuan;YE Zelan;SONG Yikang;LIN Yue;FU Hongxing(Department of Pharmacy,the First Affiliated Hospital of Ningbo University,Ningbo,Zhejiang 315010,China;Shulan International Medical College,Zhejiang Shuren College,Hangzhou,Zhejiang 310015,China;Department of Hepatobiliary and Pancreatic Surgery,the Second Affiliated Hospital,School of Medicine,Zhejiang University,Hangzhou,Zhejiang 310003,China)

机构地区：[1]宁波大学附属第一医院药学部,浙江宁波315010 [2]浙江树人学院树兰国际医学院,浙江杭州312028 [3]浙江大学医学院附属第二医院肝胆胰外科,浙江杭州310003

出　　处：《肝胆胰外科杂志》2025年第1期26-30,共5页Journal of Hepatopancreatobiliary Surgery

基　　金：宁波市自然科学基金项目(2021J248)。

摘　　要：目的基于绿色荧光蛋白(GFP)标记胰岛,建立一种简便、精准探索胰岛在大小鼠不同部位移植存活效果的方法。方法GFP腺病毒质粒转染标记分离鼠胰岛,将其移植到同系小鼠的腹腔、腹股沟皮下以及同系大鼠大网膜内,移植10 d后用便携式激发光源观察胰岛在移植部位的分布和存活情况,双硫腙(DTZ)染色液染色和倒置荧光显微镜确认胰岛,HE染色确认在移植位点的胰岛细胞团形态。结果胰岛经感染复数为100的GFP腺病毒质粒转染后再培养48 h,在荧光显微镜和便携式激发光源照射下均可见胰岛发出绿色荧光;移植200个转染GFP质粒的胰岛至小鼠腹腔、腹股沟皮下以及大鼠肠系膜内10 d后,均可发现在移植部位有发绿色荧光的胰岛组织。以小鼠腹股沟皮下移植的胰岛为例,经DTZ染色可见组织中有显红色并能表达绿色荧光的胰岛组织,经HE染色可见胰岛外形圆整。结论本研究建立了一种在动物体内探索胰岛移植存活的简便、精准方法,为初步判断该移植位点对于胰岛移植的适用性和后续进一步改进研究奠定了基础。Objective Based on green fluorescent protein(GFP)labeling of pancreatic islets,to establish a simple and accurate method,to explore the survival effect of pancreatic islet transplantation in different parts of mice and rats.Methods GFP adenovirus plasmid transfection was used to label and isolate islets from mice and rats,and transplanted islets into mice abdominal cavity,inguinal subcutaneous tissue,and rats omentum of the same strain.After 10 d of transplantation,the distribution and survival of islets at the transplantation sites were observed with a portable excitation light source.Dithizone(DTZ)staining and fluorescence microscopy were used to confirm the pancreatic islet cells,and HE staining was used to confirm the morphology of pancreatic islet cell clusters at the transplantation sites.Results After transfection with GFP adenovirus plasmid with a multiplicity of infection of 100,the pancreatic islets were cultured for 48 h,and green fluorescence was observed under fluorescence microscopy and portable excitation light source irradiation.After transplantation of 200 GFP plasmid transfected pancreatic islets into the abdominal cavity and subcutaneous inguinal region of mice,and mesentery of rats,for 10 d,green fluorescent pancreatic islet tissue was found at the sites.Taking the subcutaneous transplantation of pancreatic islet tissue in the groin of mice as an example,after frozen sectioning with 10μm thick and DTZ staining,pancreatic islet tissue with red color and green fluorescence expression could be seen in the tissue.By HE staining,the appearance of pancreatic islets was round.Conclusion This study established a simple and accurate method to explore the survival of islet transplantation in mice and rats,which laid a foundation for the preliminary judgment of the applicability of the transplantation site for islet transplantation and further improvement research.

关 键 词：绿色荧光蛋白 胰岛 小鼠 大鼠 移植部位 胰岛存活 

分 类 号：R458.5[医药卫生—治疗学]