一种小鼠胰岛肾被膜下移植方法及效果评价

A method of islet transplantation under kidney capsule in mouse and the evaluation of it＇s effect

目的 介绍一种简易的小鼠胰岛肾被膜下移植方法,并对移植后小鼠血糖、糖耐量和组织学进行评价.方法 采用BALB/c小鼠,胰岛采用小鼠逆行胆总管灌注胶原酶和Histopaque密度梯度液纯化,经手工挑选和简单培养之后,采用注射器结合电泳点样吸头将胰岛推注移植至糖尿病小鼠的肾被膜下,定期检测小鼠血糖、糖耐量试验,并于第10天取移植胰岛的肾,对肾被膜下胰岛进行苏木素-伊红（HE）和免疫荧光染色.结果 同时消化4只小鼠可得胰岛的数量为（1 063±122）个,当量为1 259.72±107.19,胰岛移植后小鼠血糖均在24 h内降至正常水平,糖耐量结果显示移植后小鼠的血糖值显著低于糖尿病小鼠（P＜0.01）,免疫荧光染色结果显示移植后胰岛活性良好[（92.8±2.6）％].结论 本实验方法操作简易,移植后胰岛活性良好,为进一步进行胰岛移植相关的体内外药物筛选等研究奠定了基础.

Objective In this paper,we introduced a simple and easy way to do islet transplantation under kidney capsule in mouse,and the effect evaluation of it＇ s blood glucose,oral glucose tolerance and histological stain after transplantation.Methods BALB/c mice are used in the experiment.The islets were harvested by reversely perfusion of collagenase V into common bile duct and purification by Histopaque density gradient followed by manual picking.After a simple culture,the islets were transplanted under the kidney capsule of diabetic BALB/c mice using a gel loading tip.Then the blood glucose and the oral glucose tolerance test were tested regularly.The kidney was cut at the 10th day after the islet transplantation.The HE and immunofluorescence staining of islet under kidney capsule were also done to evaluate the islet morphology and function.Results We get 1 063 ± 122 islets,or 1 259.72 ± 107.19 IEQ from four mice at one time.The level of blood glusoce turned down to normal in 24 h after the islet transplantation.Oral glucose tolerance test （OGTY） results showed that the blood glucose levels of mice after islet transplantation were significantly lower than those of diabetes （P 〈 0.01）.And the results of immunofluorescence staining showed that the activity of islets was good [（92.8 ± 2.6） %].Conclusion The method of the experiment is simple,and the islet activity is good after transplantation,which make a good foundation for the further research on drug screening for islet transplantation.