三维微重力培养大鼠胰岛细胞的实验研究

Experimental study on islet cells in rats under condition of three-dimensional microgravity

目的 应用三维微重力组织培养对大鼠胰岛细胞的存活率及分泌功能进行研究。方法将大鼠胰岛细胞分离消化后分别进行二维普通培养 (Ⅰ组 )及三维微重力条件培养 (Ⅱ组 ) ,应用双硫腙 (DTZ)染色法 ,对胰岛细胞进行特异性染色鉴定 ;采用AO PI双染色法对两组培养 3d、7d、14d的胰岛细胞存活率进行检测 ;应用放射免疫法检测两种不同培养方法培养液中胰岛素分泌水平。结果 DTZ染色后胰岛呈桔红色 ,清晰可见。培养 7d和 14d时 ,Ⅱ组胰岛细胞存活率分别为(0 90 0 0± 0 0 10 7) %和 (0 80 38± 0 0 0 92 ) % ,均明显高于Ⅰ组 (P <0 0 1)。胰岛素测定结果表明 ,培养 7d时 ,Ⅱ组胰岛素水平为 (70 875± 0 31)mU/L ,Ⅰ组胰岛素水平为 (41 2 4 6± 0 35 )mU/L ,二者差异有显著意义 (P <0 0 1)。在培养的 14d、2 1d和 30d时 ,Ⅱ组胰岛素的分泌水平也均高于Ⅰ组(P <0 0 1)。结论 三维微重力培养组的胰岛细胞存活率及胰岛素分泌功能都优于二维普通培养组。

Objective To study the survival rate and secretory function of islet cells in rats under condition of three-dimensional microgravity. Methods Isolated islet cells were assigned to flask-cultured or bioreactor-cultured. Survival rate of islets cultured for days 3?7?14 in stationary flasks or microgravity bioreactors was measured by AO-PI double-staining. Cultured islets were identified by dithizone (DTZ) staining,and insulin contents of different culture liquids were measured by radioimmunoassay. Results Pancreatic islets stained nacarat with DTZ were easily visualized. When islet cells were cultured for 7 days and 14 days,survival rate of bioreactor-cultured islets was (0.9000±0.0107)% and 0.8038%±0.0092% and higher than flask-cultured islets ( P <0.01). Insulin level of bioreactor-cultured islets is (70.875±0.31) mμ/L on the cultured 7days while flask-cultured islets is (41.246±0.35) mμ/L. There was statistically significant difference of insulin production between the two groups ( P <0.01). Bioreactor-cultured islet contents were higher than flask-cultured on the cultured 14 ?21 and 30 days( P <0.01). Conclusions Islet cells survival rate and secretory function revealed that bioreactor-cultured islets functioned better compared with flask-cultured islets.