高浓度大鼠肝实质细胞与非实质细胞共培养的细胞功能研究  

Study on Specific Hepatocellular Function of High Density Rat Hepatocytes Co-cultured with Nonparenchymal Liver Cells

作　　者：舒桂明[1] 宋继昌[1] 杜智[1] 李涛[1] 张金卷[1] 付丽[1] 吕文岚[1] 崔润海[1] 毕海英[1] 

机构地区：[1]天津市第三中心医院肝胆外科,天津市肝胆疾病研究所,卫生部人工细胞工程技术中心,300170

出　　处：《天津医药》2002年第6期353-356,I004,共5页Tianjin Medical Journal

基　　金：天津市科委发展计划项目(项目编号:983113611)

摘　　要：目的:高浓度肝实质细胞与非实质细胞原代共培养,研究其功能活性。方法:应用原位胶原酶灌流法分离大鼠肝细胞,获得有活性的肝细胞,并应用高浓度实质和非实质肝细胞共培养的方法原代培养（共培养组）,并以微囊肝细胞培养为对照组（微囊组）进行了比较。结果:两组均维持白蛋白分泌、尿素合成功能7天;共培养组的白蛋白分泌与微囊组一样为下降趋势,共培养组从（0.870±0.102）降至（0.492±0.040）g·L-1·10-6cells·24h-1,微囊组从（1.147±0.099）降至（0.375±0.012）g·L-1·10-6cells·24h-1;共培养组的肝细胞第4天后维持在一个较稳定的水平,而微囊组肝细胞前3天明显高于共培养组,而后两天显著低于共培养组（P<0.05）。共培养组的尿素合成功能,由（4.50±0.56）降至（4.37±0.19）μmol·L-1·10-6cells·90min-1,微囊组由（5.42±0.81）降至（3.60±0.33）μmol·L-1·10-6cells·90min-1,前3天微囊组高于共培养组,后2天共培养组明显高于微囊组（P<0.05）。共培养2～7天肝细胞的对氨基苯甲酸（PABA）浓度稳定在 7.2～9.9mg/L·10-6cells·24h-1。结论:高浓度肝实质细胞与非实质细胞共培养,可使肝细胞的特异性功能维持7d,而且较微囊肝细胞更适合应用于中空纤维舱型生物人工肝。Objective:To study the culturing of primarily high density of hepatocyes with ninparenchymal liver cells and specific hepatocellular function. Methods: Rat liver cells were isolated by perfusing collagenase in situ, and high density of hepato-cytes (107 cells/mL) were co-cultured primarily with nonparenchymal liver cells (co-culture group), which specific functions were measured and contrasted with encapsulated hepatocytes(encapsule group) . Results: The levels of albumin section and urea synthesis were maintained in 7 days. Albumin level descended from (0.870 ± 0.102) to (0.492 ± 0.040)g-L 1-10-6cells-24h-1 in co-culture group and from (1.147 ± 0.099) to (0.375 ± 0.012)g-L -1 - 10 6cells-24h-1 in encapsule group; level of albumin was significantly higher in the former 3 days in encapsule group, whereas significantly higher in the later 2 days in co-culture group. Urea synthesis descended from (4.50 ± 0.56) to (4.37 ± 0.19)μmol- L-1-10-6 cells-90min-1 in co-culture group and from (5.42 ±0.81) to (3.60±0.33)μmol-L-1-10-6cells-90min-1 in encapsule group; urea concentration was significantly higher in the former 3 days in encapsule group whereas significantly higher in the later 2 days in co-culture group. Para-aminobenzoic acid(PABA) concentration in co-culture group ranged from 7.2 to 9.9mg-L-1-10-6cells-24h-1. Conclusion: Culturing of high density hepatocytes with nonparenchymal liver cells can preserve specific hepatocellular function for 7 days, which is more suitable to bioartificial liver systems as biological component than encapsulated hepatocytes.

关 键 词：高浓度 大鼠 肝实质细胞 非实质细胞 共培养 细胞功能 研究 

分 类 号：R333.4[医药卫生—人体生理学]