Sheil A G, Sun J, Mears D C, Waring M, Woodman K, Johnston B, Horvat M, Watson K J, Koutalistras N, Wang L S
Australian National Liver Transplantation Unit, Royal Prince Alfred Hospital, Sydney, New South Wales, Australia.
Aust N Z J Surg. 1996 Aug;66(8):547-52. doi: 10.1111/j.1445-2197.1996.tb00807.x.
This study describes the pre-clinical trials of an extracorporeal bioartificial liver support system (BALSS). It includes the biochemical changes which occur in the plasma and blood of pigs with devascularized livers when the plasma is treated in a BALSS, and the testing of the system for presence or absence of infective agents, pyrogens and for toxicity.
Hepatic cells were prepared from littermate juvenile white landrace pigs with a double-step collagenase digest technique. The cell preparations were incubated with collagen-coated dextran microspheres (CDM) for 3 h and the medium was tested to determine cellular metabolic activity. Incubation continued for a further 20 h during which the hepatic cells attach to the CDM. The CDM-attached cells were inoculated into a hollow fibre bioreactor which was part of an extracorporeal liver support system.
Hepatic cell content of the bioreactor was 6 x 10(9) +/- 3 x 10(8) cells, equivalent to those present in half a pig's liver. The system was tested in a controlled trial with the plasma of pigs with fulminant hepatic failure (FHF) due to devascularized livers. When plasma from FHF pigs was circulated through the device there was significantly less of an increase in the accumulation of ammonia, lactate and most amino acids when hepatic cells were included in the circuit compared with those in control experiments when they were excluded. Similar changes occurred in procine blood. There were few infections diagnosed and an absence of pyrogens, endotoxins and toxicity in the bioreactor contents or in the terminating reservoir or animal blood samples.
We believe that the results, demonstrating function of the porcine hepatic cells in the circuit, together with low risks, justify a clinical trial of use of the BALSS in Australia.
本研究描述了一种体外生物人工肝支持系统（BALSS）的临床前试验。它包括当血浆在BALSS中处理时，去血管化肝脏的猪的血浆和血液中发生的生化变化，以及该系统是否存在感染因子、热原和毒性的测试。
采用两步胶原酶消化技术从同窝幼龄白色长白猪制备肝细胞。将细胞制剂与胶原包被的葡聚糖微球（CDM）孵育3小时，并检测培养基以确定细胞代谢活性。孵育持续另外20小时，在此期间肝细胞附着于CDM。将附着有CDM的细胞接种到作为体外肝支持系统一部分的中空纤维生物反应器中。
生物反应器中的肝细胞含量为6×10⁹±3×10⁸个细胞，相当于半只猪肝中的细胞数量。该系统在一项对照试验中进行了测试，使用的是因去血管化肝脏导致暴发性肝衰竭（FHF）的猪的血浆。当FHF猪的血浆通过该装置循环时，与排除肝细胞的对照实验相比，当回路中包含肝细胞时，氨、乳酸和大多数氨基酸的积累增加明显较少。猪血液中也发生了类似变化。在生物反应器内容物、终末储液器或动物血液样本中几乎没有诊断出感染，也没有热原、内毒素和毒性。
我们认为，这些结果证明了回路中猪肝细胞的功能，同时风险较低，为在澳大利亚对BALSS进行临床试验提供了依据。
