CAJ | 학술논문

目的探讨胰腺干细胞体外定向分化潜能,评价诱导分化后细胞同种移植对糖尿病的治疗效果.方法体外分离、纯化成体Wistar大鼠胰腺干细胞,运用荧光免疫染色法对胰腺干细胞表面进行鉴定,然后在肝细胞生长因子、尼克酰胺等共刺激下诱导细胞定向分化.双硫腙(DTZ)染色对诱导后细胞进行胰岛细胞鉴定,ELISA染色光电酶标记法检测其胰岛素分泌功能.采用链脲佐菌素(STZ)腹腔注射建立Wistar大鼠糖尿病模型,将40只造模成功的大鼠随机分为胰岛细胞移植组(移植组)和安慰剂注射组(对照组),分别于移植前1 d及移植后1、2、3、4周经尾静脉采血检测血清胰岛素和血糖水平.结果本实验成功获取成体大鼠胰腺干细胞,细胞表面CK19、Pdx-1和Nestin表达均为阳性.诱导分化后的细胞DTZ染色呈棕红色,在高糖刺激下表达、分泌胰岛素.移植后4周内移植组血清胰岛素水平平均为(11.41±1.52)mU/L,血糖水平平均为(8.22±2.7)mmol/L,对照组分别为(9.30±1.56)mU/L和(12.23±3.8)mmol/L,两组相差显著(P＜0.05).结论体外分离成体胰腺干细胞可被定向诱导分化为具有胰岛素分泌功能的胰岛样细胞,同种移植后对糖尿病具有治疗作用.
목적 탐토이선간세포체외정향분화잠능,평개유도분화후세포동충이식대당뇨병적치료효과.방법 체외분리、순화성체Wistar대서이선간세포,운용형광면역염색법대이선간세포표면진행감정,연후재간세포생장인자、니극선알등공자격하유도세포정향분화.쌍류종(DTZ)염색대유도후세포진행이도세포감정,ELISA염색광전매표기법검측기이도소분비공능.채용련뇨좌균소(STZ)복강주사건립Wistar대서당뇨병모형,장40지조모성공적대서수궤분위이도세포이식조(이식조)화안위제주사조(대조조),분별우이식전1 d급이식후1、2、3、4주경미정맥채혈검측혈청이도소화혈당수평.결과 본실험성공획취성체대서이선간세포,세포표면CK19、Pdx-1화Nestin표체균위양성.유도분화후적세포DTZ염색정종홍색,재고당자격하표체、분비이도소.이식후4주내이식조혈청이도소수평평균위(11.41±1.52)mU/L,혈당수평평균위(8.22±2.7)mmol/L,대조조분별위(9.30±1.56)mU/L화(12.23±3.8)mmol/L,량조상차현저(P＜0.05).결론 체외분리성체이선간세포가피정향유도분화위구유이도소분비공능적이도양세포,동충이식후대당뇨병구유치료작용.
Objective To investigate the potential of pancreatic stem cells (PSCs) directed differentiation in vitro, and to evaluate the effects of differentiated PSCs allograft on the treatment of diabetes.Methods The PSCs of adult Wistar rats were separated and purified in vitro. The surface of PSCs was determined by immunofluorescence staining, and then it was stimulated by hepatocyte growth factor (HGF) and nicotinamide to induce directed differentiation. Dithizone dyeing was used to determine the islet-like cells after induction, and ELISA staining method was used to detect the insulin levels. Streptozotocin peritoneal injection was used to induce the diabetic rat mode. 40 rats were randomly allocated into pancreatic islet cells allograft group (experiment group) and placebo group. The serum insulin and glucose levels 1 d before transplantation and 1, 2, 3, 4 week after transplantation were measured. Results PSCs of adult Wistar rats were successfully obtained, and the expression of CK19, Pdx-1 and Nestin on cell surface was positive. Dithizone dyeing for directed differentiation cells showed brownish red color. The cells could express and secrete insulin after hyperglycaemia stimulation. The serum insulin and glucose levels 4 week after transplantation were (11.41 ±1.52) mU/L and (8.22 ± 2.7) mmol/L, which were (9.30 ± 1.56) mU/L and (12.23 ± 3.8) mmol/L in the placebo group, and difference was statistically significant (P＜0.05). Conclusions PSCs can be induced and directed differentiated in vitro into islet-like clusters with insulin secretion function. And its allograft has the potential for the treatment of diabetes.