人脐静脉间充质干细胞向胰岛β样细胞诱导分化的实验研究  

Differentiation of mesenchymal stem cells derived from human umbilical vein into islet β-like cells in vitro

作　　者：李德华[1] 单伟[1] 韩静[2] 赵宝东[1] 秦书俭[1] 

机构地区：[1]辽宁医学院解剖学教研室 [2]辽宁医学院附属三院内科,辽宁锦州121001

出　　处：《中国临床解剖学杂志》2007年第4期424-426,共3页Chinese Journal of Clinical Anatomy

基　　金：辽宁省自然科学基金(2004C039)

摘　　要：目的:探讨人脐静脉内皮及内皮下间充质干细胞向胰岛β样细胞分化的可行性。方法:采用胶原酶消化分离人脐静脉内皮及内皮下细胞进行贴壁培养;传代培养后,用高浓度葡萄糖(25mmol/l)培养液DMEM(含10%FBS)以及bFGF(basic Fibroblast Growth Factor)和尼克酰胺诱导脐静脉间充质干细胞向胰岛β样细胞分化。倒置显微镜下观察间充质干细胞诱导后形态变化,观察是否形成细胞团;用胰岛β细胞特异双硫腙染色鉴定诱导后细胞内是否含有高浓度游离锌离子;免疫细胞化学(Envision法)鉴定诱导后细胞是否分泌胰岛素。结果:第2代脐静脉间充质干细胞经过高糖诱导大约10天后,形成细胞团;呈双硫腙染色阳性;免疫细胞化学表明诱导后细胞内的细胞胰岛素染色阳性。结论:人脐静脉内皮及内皮下分离出的间充质干细胞在体外可以定向诱导分化为胰岛β样细胞,这种胰岛β样细胞具有胰岛素的分泌功能。Objective： To isolate mesenchmal stem cells （MSCs） from endodermis and subendodermis of human umbilical vein and induce them into islet β-like cells. Methods： By the digestion of collagen Ⅰ , MSCs were isolated from endodermis and subendodermis of human umbilical vein by adhesive screening method and induced by adding high glucose, bFGF （basic Fibroblast Growth Factor） and nicotamine into culture medium. Cells were stained with dithizone （DTZ）which is a zinc-chelating agent known to selectively stain pancreatic β cells because of their high zinc content, while insulin content in the treated cells was examined by immunocytochemistry. Results： Under induction conditions as described above, cultures formed homogeneous spindle-shaped cells after 2 passages. After approximately 10 days of treaanent, cells formed DTZ-stained cell clusters. Immunocytochemistry also confirmed that these cell clusters were positive for insulin. Conclusions： MSCs derived from endodermis and subendodermis of human umbilical vein can be induced into islet β-cells in vitro. These islet β-cells have secretion function of insulin. MSCs originated from human umbilical veinmay be an ideal seed cells for treating type Ⅰ diabetes.

关 键 词：人脐静脉 间充质干细胞 体外分化 胰岛 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R458.8[医药卫生—基础医学]