中文摘要：

背景：微载体培养技术作为一项体外高浓度细胞培养技术，近年来已在肝细胞的体外培养中得到应用。目的：对壳聚糖球形多孔微载体培养的人肝细胞L-02进行定时的形态学观察。方法：以自制的壳聚糖球形多孔微载体样本为支架来培养人肝细胞 L-02设为实验组；无壳聚糖球形多孔微载体支持下人肝细胞的培养设为对照组。对两组细胞进行定时的细胞计数，对实验组进行形态学观察，包括倒置相差生物显微镜观察和扫描电子显微镜观察。结果与结论：两组培养的细胞数量均呈现前3 d增长，在培养第3天细胞数量达到最高值；实验组3个样本培养的细胞数明显高于对照组无微载体培养的细胞数量（P0.05）。倒置相差生物显微镜下动态观察，可见前3 d微载体表面黏附生长的肝细胞则逐渐增多，培养第3天可见大部分微载体表面有许多肝细胞黏附成团，总的存活率均在90%以上，且肝细胞保持着良好的形态学结构；扫描电子显微镜观察，微载体表面、切面和内部均可看到有许多球状肝细胞紧密黏附。结果说明，以自制的壳聚糖球形多孔微载体作为一种支架，在体外三维环境下可以进行高浓度细胞培养。

英文摘要：

Abstract BACKGROUND： Recently, microcarrier culture as a high concentration cel culture technology has been used in hepatocyte culturein vitro. OBJECTIVE：To periodicaly observe the morphological changes of human hepatocyte L-02 cultured by the spherical porous chitosan microcarrier. METHODS：The self-made spherical porous chitosan microcarrier sample as a support to culture human hepatocytes L-02 acted as experimental group; non-spherical porous chitosan microcarrier as a support to culture human hepatocytes L-02 as control group. Cels in two groups underwent cel count at regular time, and the morphological changes were observed in the experimental group, including inverted phase contrast biological microscope observation and scanning electron microscope observation. RESULTS AND CONCLUSION：The quantity of cultured cels in the two groups was increased in the first 3 days and reached the peak at the 3rd day. The cel quantity in the experiment group was obviously higher than that in the control group （P 0.05）. The quantity of hepatocytes adhered to the microcarrier surface was gradualy increased in the former 3 days under the inverted phase contrast biological microscope. There were lots of cel clusters on the surface of a greater part of microcarriers, with the total survival rate of more than 90%, and hepatocytes kept a good morphological structure. Under the scanning electron microscope, lots of hepatocytes adhered tightly to each other on the surface and section of microcarrier as wel as inside the microcarrier. It is indicated that the self-made spherical porous chitosan microcarrier as a support in three-dimensional environment can undergo cel culture with high concentration.