Timed morphological changes of human hepatocytes L-02 cultured at high density by the support of spherical porous chitosan microcarriers / 中国组织工程研究

ABSTRACT

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.