Development of bioartificial liver support utilizing hepatocyte spheroids and its evaluation on pigs with liver failure.

利用肝细胞球体开发生物人工肝支持及其对肝功能衰竭猪的评估。

基本信息

批准号：
07457595
负责人：
KOIDE Norio
金额：
$ 0.9万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

项目摘要

A bio-artificial liver support utilizing hepatocyte multicellular spheroids (spheroid) were developed. Isolated rat or porcine hepatocytes were allowd to form spheroids in the primary culture using chemically defined medium and culture substrates of proteoglycans and their analogs. For the large scale production of spheroid porcine hepatocytes were cultured in the multilayr-plates pre-coated with neo-proteoglycan that was chemically synthesized with chondroitin sulfate and phosphatidyl ethanolamine. Functional level of the spheroids were evaluated with the activities of p-450s, albumin production and urea production. A bio-artificial liver support system was consisted with a closed circuit to which a bioreactor chambercontaining hapatocyte spheroids was inserted. The closed circuit was equipped with a regulator unit that automatically regulated the levels of dissolved oxygen and pH in the circulating medium. A hollow fiber cartridge for plasma exchange or a spouted bed chamber were used as the bioreactor chamber. The performance of the bio-artificial liver support system was evaluated on pigs with acute liver failure experimentally induced by the administration of D-galactosamine. The reductio of serum ammonia level was observed in all of the pigs treated with bio-artificial liver support system. However no significant improvement of the survival rate was seen, though the elongation of survival time was in some pigs. These result obtained with hepatocyte spheroid equivalent to about 10% of liver cell volume in vivo.

开发了利用肝细胞多细胞球体（球体）的生物人工肝支持。使用化学成分确定的培养基和蛋白聚糖及其类似物的培养基质，使分离的大鼠或猪肝细胞在原代培养物中形成球状体。为了大规模生产球状猪肝细胞，在预先涂有新蛋白聚糖的多层板中培养，所述新蛋白聚糖是用硫酸软骨素和磷脂酰乙醇胺化学合成的。通过p-450s的活性、白蛋白产生和尿素产生来评估球体的功能水平。生物人工肝支持系统由闭路组成，其中插入了含有肝细胞球体的生物反应器室。闭合回路配备了调节装置，可自动调节循环介质中的溶解氧和 pH 值。用于血浆交换的中空纤维盒或喷射床室用作生物反应器室。在通过给予 D-半乳糖胺实验诱导的急性肝衰竭猪身上评估了生物人工肝支持系统的性能。所有接受生物人工肝支持系统治疗的猪均观察到血清氨水平降低。然而，虽然一些猪的存活时间有所延长，但存活率没有显着提高。这些结果是用相当于体内肝细胞体积约10%的肝细胞球体获得的。