摘要
目的建立一种有效的原代猪肝细胞长期培养体系．为人工肝生物材料提供生存环境．方法把经二步胶原酶灌注法分离的猪肝细胞按３×１０５分别接种于无血清培养基、１００ｍＬ／Ｌ胎牛血清培养基、１００ｍＬ／Ｌ的猪门静脉血清培养基中．光镜下观察各体系中肝细胞形态变化过程．采用Ｂｅｃｋｍａｎ全自动生化分析仪检测各培养体系不同时间培养上清中Ａｌｂｕｍｉｎ，Ｕｒｅａ的含量．结果原代猪肝细胞在三种不同培养体系中存活时间分别是：无血清培养基４ｄ～５ｄ，胎牛血清培养基３５ｄ～３７ｄ，猪门静脉血清培养基５７ｄ～５８ｄ．存活的肝细胞功能活性除无血清培养组不明显外，胎牛血清培养组可维持４ｗｋ左右，猪门静脉血清培养组则可维持８ｗｋ左右结论门静脉血清培养体系是原代猪肝细胞较为理想的生存环境．
AIM To establish an effective primary porcine hepatocytes culturing system for the biological material of bioartificial liver(BAL). METHODS Porcine hepatocytes were harvested by two step perfusion collagenase method. Individually, cells were plated at a concentration of 3×10 5 in serum free DMEM culture medium, DMEM culture medium supplemented with 10% fetal calf serum or 100mL/L porcine portal vein serum. Changes in cell spreading with time in culture were determined by taking phase contrast micrographs of cells. The concentration of albumin and urea were measured. RESULTS The primary porcine hepatocytes viability and specific function were maintained in serum free culture system for 7 days, in FCS system for 35-37 days,and in porcine portal vein serum system for over 50 days. CONCLUSION The culture medium supplemented with porcine portal vein serum is an ideal circumstance for maintaining porcine hepatocytes viability in vitro .
出处
《世界华人消化杂志》
CAS
1999年第3期206-209,共4页
World Chinese Journal of Digestology
基金
国家自然科学基金
关键词
人工肝
肝细胞
细胞培养
artificial liver
hepatocytes
cell, cultured
