大鼠肝细胞与睾丸支持细胞混合共微囊化的体外功能研究  

The function of mixed-microencapsulated rat hepatocytes and testicular Sertoli cells in vitro

作　　者：郑明华[1] 陈永平[1] 林海龙[1] 潘亮亮[2] 许烂漫[1] 黄瑜[1] 马海龙[1] 潘珍珍[1] 

机构地区：[1]温州医学院附属第一医院感染内科,325000 [2]温州医学院基础医学院

出　　处：《中华传染病杂志》2007年第9期513-516,共4页Chinese Journal of Infectious Diseases

基　　金：浙江省2004年度引进国外技术;管理人才专项资助项目(Z2004GY031);温州市科技计划资助项目(Y2004A011)

摘　　要：目的观察大鼠肝细胞与睾丸支持细胞混合共微囊化对肝细胞生物学活性的支持作用。方法利用微囊发生器制备含肝细胞、睾丸支持细胞及两者混合细胞的微囊,根据微囊内包裹细胞种类的不同,分为微囊化肝细胞组,微囊化睾丸支持细胞组,肝细胞、睾丸支持细胞分别微囊后共同培养组和肝细胞、睾丸支持细胞混合共微囊组。观察囊内细胞形态,体外培养测定培养液中Alb与尿素分泌,判断各组囊内肝细胞活性和功能。样本比较采用重复测量数据的多元方差分析。结果体外培养96 h时,光学显微镜下见微囊呈球形,表面光滑,细胞均匀、散在分布于微囊内,囊内肝细胞、睾丸支持细胞呈圆形,未贴壁。微囊化睾丸支持细胞组培养上清液中无法测及Alb与尿素。与微囊化肝细胞组相比,肝细胞、睾丸支持细胞分别微囊后共同培养组与肝细胞、睾丸支持细胞混合共微囊组肝细胞的存活时间延长,培养上清液中Alb(F=217.56,P<0.01)和尿素(F=232.72,P<0.01)明显增加。混合细胞共微囊组于培养第7天Alb与尿素含量最高,分别为(2.80±0.11)g/L、(1.92±0.10)μmol/L,而其他两组均于培养第3天Alb与尿素含量达到最高,分别为(2.48±0.08) g/L、(1.17±0.08)μmol/L和(2.27±0.10)g/L、(1.37±0.05)μmol/L。相对于混合细胞共微囊组,肝细胞、睾丸支持细胞分别微囊后共同培养组在培养中后期Alb、尿素含量下降趋势明显。结论肝细胞与睾丸支持细胞混合共微囊化能明显延长囊内肝细胞的寿命,维持肝细胞形态和功能。Objective To investigate the supportive effects of mixed microcapsules of rat hepatocytes and testicular Sertoli cells on liver function. Methods Microcapsule containing hepatocytes and（or） testicular Sertoli cells was prepared by microcapsule generator, According to the type of microencapsulated cells, the microcapsules were divided into four groups： microencapsulated hepatocytes alone （group Ⅰ ）, microencapsulated Sertoli cells alone （group Ⅱ ）, mixture of microencapsulated hepatocytes and microencapsulated Sertoli cells （group Ⅲ ）, and mixed-microencapsulated hepatocytes and Sertoli cells （group Ⅳ ）. Morphology of the microencapsulated cells was observed by phase contrast microscopy. Supernatant was collected regularly to detect the secretion of albumin and urea to evaluate the activities of hepatocytes in different groups. The repeated measurement of MANOVA was performed for the data analysis. Results After 96 hours encapsulation, hepatocytes and Sertoli cells were dispersed evenly in the microencapsulates, which were sphere with smooth surface and non-adherent. Albumin and urea in the supernatant of group Ⅱ were undetectable. Compared with group Ⅰ, cells in group Ⅲ and Ⅳ survived much longer and the levels of albumin and urea in the supernatant of group Ⅲ and Ⅳ increased significantly （F 217.56, P〈0.01; F= 232.72, P〈0.01, respectively）. The levels of albumin and urea in group Ⅳ peaked on day 7 which were （2.80±0.11） g/L and （1.92±0. 10） μmol/L, respectively, while in group Ⅰ and Ⅲ, the levels peaked on day 3 [albumin：（2.27±0. 10） g/L, urea：（1. 37±0.05） μmol/L for group Ⅰ albumin：（2.48±0.08） g/L, urea： （1.47±0.08） μmol/L for group Ⅲ, respectively3. Compared with group IV, the levels of albumin and urea in group Ⅲ decreased significantly in the late phase of microencapsulation. Conclusions The hepatocytes and testicular Sertoli cells mixed-microencapsulation can significantly improve the survival time and mai

关 键 词：肝细胞 塞尔托利细胞 微囊 睾丸 大鼠 

分 类 号：R657.3[医药卫生—外科学]