壳聚糖微载体的制备及原代大鼠肝细胞培养

PREPARATION OF CROSSLINKED CHITOSAN MICROCARRIER AND ITS USE IN CULTURE OF PRIMARY RAT HEPATOCYTE

目的 探讨制备壳聚糖微载体在原代大鼠肝细胞培养中的应用效果。方法 利用甲苯 -四氯化碳作有机分散介质 ,戊二醛作交联剂 ,通过反相悬浮交联制备微米级的壳聚糖微载体。用其进行原代大鼠肝细胞培养 ,利用相差显微镜和扫描电镜对细胞形态进行观察 ,测定细胞的代谢活性。结果 通过对壳聚糖浓度和戊二醛用量等反应条件的优化 ,制成了性能优良的微载体。肝细胞在壳聚糖微载体上保持良好的球形状态 ,白蛋白分泌可维持 7天以上 ,最高分泌量达到 2 6 .7μg/2 4 h/ml。结论 壳聚糖微载体是一种优良的肝细胞培养支架。

Objective To prepare chitosan microcarriers and to use it to cultivate rat primary hepatocytes. Methods The crosslinked chitosan microcarrier was prepared by the reaction of glutaraldehyde with chitosan. Various factors that influence the preparation were studied and the reaction conditions were optimized. Rat primary hepatocytes cultured on chitosan microcarrier were observed by using phase contrast microscope and scanning electron microscope. Results Chitosan microcarriers with good properties could be prepared by adjusting the concentration of chitosan solution and the quantity of glutaraldehyde. Rat hepatocytes cultured on chitosan microcarriers retained the spherical shape as they have in vivo. And albumin secretion can last over one week. The highest albumin secretion rate reached 26.7μg/24h/ml. Conclusion Chitosan microcarriers is a promising scaffold for hepatocyte attachment, which can be used in bioartificial liver support system.