摘要
目的为体外生物人工肝支持系统构建一种新型的多层平板型生物人工肝细胞反应器，从而提高反应器中细胞的活性及功能。方法自主设计组装一种新型的多层平板型生物反应器，平板表面采用静电纺丝纺上一层壳聚糖纳米纤维膜，新鲜分离的原代猪肝细胞接种于纳米纤维支架表面，用培养液进行循环式培养。定期检测反应器中细胞的活性及功能，并观察细胞与纳米材料结合情况。结果反应器中的细胞能与纳米支架材料很好的结合，每12h分泌的白蛋白的量维持在100～139ug，尿素合成量为5．11～7．76mg／dl，而氯化铵清除率则保持在44—60umol／h。培养过程中，仅有少量的酶（AST、LDH）漏出。结论初步构建的新型多层平板型生物反应器，符合生物人工肝基本的要求。其内培养的细胞能保持良好的活性，并具有相应的功能。
Objective To develop a novel multi-layer flat plate bioreactor based on nanofiber scaffolds, so as to improve the viability and functions of hepatocytes in the bioreaetor. Methods The new bioreactor we designed consisted of 50 layers of stacked flat plates, on which chitosan nanofiber scaffold was electrospinned for cell immobilization and aggregation. Porcine hepatocytes were cultured on the nanofiber scaffolds. Parameters for hepatocyte-specific function and enzyme leakage were determined. Results Hepatocytes adhered onto the nanofiber scaffold tightly, and secreted albumin at a level of 100-139ug/12 h, the urea synthesis was 5.11-7.76 mg/dl, and the ammonia elimination was 44-60 umol/h. During the experiment, only a little of AST and LDH were released from the cultured hepatocytes into the medium. Conclusion The new bioreactor we designed met the needs of bioartificial liver very well, and the cells in the bioreaetor could maintain their viability and keep the liver-specific functions.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2009年第9期1150-1152,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（30772129）
