微囊化培养对肝细胞增殖生长和功能表达的影响  

Effect of microencapsulated culture on the proliferation and functional expression of hepatocytes

作　　者：黄晓波[1] 张英[1] 王为[1] 孙广炜[1] 朱静[1] 谢红国[1] 马小军[1] 

机构地区：[1]中国科学院大连化学物理研究所生物医用材料工程组,辽宁省大连市116023

出　　处：《中国组织工程研究与临床康复》2010年第29期5329-5333,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家科技部"973"项目(2005CB522702);国家自然科学基金重点项目(20736006);国家863重大项目(2006AA02A140)~~

摘　　要：背景:前期研究证实,肝细胞在微囊化培养过程中结构形态发生变化,细胞的骨架会发生重排。目的:在前期研究基础上,进一步考察微囊化培养体系对细胞生长和功能表达的影响。方法:采用海藻酸钠-多聚赖氨酸-海藻酸钠微囊化人肝癌细胞系HepG2细胞,通过显微观察、苏木精-伊红染色、MTT实验、Realtime RT-PCR与Elisa实验,检测微囊内细胞的生长形态、细胞团的形态结构、细胞的活性、以及细胞的功能表达情况。结果与结论:HepG2细胞在微囊内聚集成团,以三维方式生长,细胞间连接紧密,与传统的平面培养相比,微囊化细胞的生长速率减慢,但微囊化细胞的功能基因表达水平及白蛋白的分泌量增高,说明海藻酸钠-多聚赖氨酸-海藻酸钠微胶囊所提供的微环境有利于肝细胞类组织的体外构建。结果也提示海藻酸钠-多聚赖氨酸-海藻酸钠微胶囊能够实现肝细胞的体外类组织化培养,有望成为一种更具前景的三维培养模式,应用于癌症治疗、高通量药物筛选以及肝组织工程的研究。BACKGROUND：The previous experiments have proved that,the configuration of hepatocarcinoma cells（HepG2） was changed during microencapsulated culture and the cellular skeleton was also rearranged.OBJECTIVE：Based on the previous study,this study was aimed to further investigate the effect of microencapsulated culture on the proliferation and function expression of HepG2 cells.METHODS：Human HepG2 cell lines were encapsulated in alginate-poly-L-lysine-alginate microcapsules.The configuration and microstructure of cellular spheroids,cellular proliferation and function expression were detected through microscopy and hematoxylin-eosin staining,MTT assay,Realtime RT-PCR and Elisa assay,respectively.RESULTS AND CONCLUSION：Microencapsulated HepG2 cells aggregated into clumps with tight junctions and proliferated in three-dimensional manner.Compared with the monolayer culture,the rate of cellular proliferation in microcapsules was reduced.However,the gene expression related with liver function and albumin secretion were increased in the microencapsulated HepG2 cells.The microenvironment provided by alginate-poly-L-lysine-alginate microcapsules can contribute to in vitro construction of HepG2 cells.The alginate-poly-L-lysine-alginate microcapsules can achieve in vitro culture of hepatocytes,it is a potential pattern of three-dimensional culture,and can be used in cancer therapy,high-flux drug screening and liver tissue engineering study.

关 键 词：微胶囊 三维培养 细胞聚集体 人肝癌细胞株 生物材料与药物控释 

分 类 号：R318[医药卫生—生物医学工程]