Hossein Aghdaie M, Azarpira N, Shamsaeefar A, Motazedian N, Kaviani M, Esfandiari E, Golbabapour S, Nikeghbalian S, Kazemi K, Salahi H, Malek-Hosseini S A, Geramizadeh B
Transplant Research Center, Shiraz University of Medical Sciences.
Department of Hepatobiliary Surgery, Shiraz University of Medical Sciences.
Int J Organ Transplant Med. 2020;11(1):15-25.
Hepatocyte transplantation using isolated human hepatocytes is an alternative source that can be used for the treatment of metabolic diseases and acute liver failure as a time bridge to liver transplantation. These cells can also be used for bioartificial liver systems and study of drug toxicity.
To determine which cold preservation solution is better maintain the liver function.
We prepared 4 cold preservation solutions made of different combination of antioxidants, chelating, membrane protective, and anti-apoptotic agents as well as inhibitor of cyclophilin D. For hepatocyte isolation, we used livers obtained from unused deceased donor livers and the liver of patients with Crigler-Najjar syndrome who were candidates of partial liver transplantation. After culture and cold preservation, the level of albumin, and urea production were measured as indices of liver functionality.
We found that albumin production significantly decreased after cold preservation in solution 1. There was no significant difference in urea production after cold preservation in solution 1 compared with control 24 h. No significant differences in albumin production were found after cold storage in solution 2 and solution 4 compared with control 24 h. Urea production significantly decreased after cold storage in solutions 2 and 4 compared with control 24 h. As a whole albumin and urea production were significantly decreased after cold preservation. Although albumin and urea production were decreased after cold preservation, but the results of albumin production of two solutions were not significantly different from that of the control group (p=0.109 and 0.951).
Cold preservation of cultured human hepatocytes in solution 2 and solution 4 could maintain the function of albumin production better than other cold preservation solutions in our experiments; solution 1 was more effective on urea production of cultured human hepatocytes at 4 °C for 24 h. To determine if these hepatocytes are suitable candidates for transplantation, further studies should be performed.
使用分离的人肝细胞进行肝细胞移植是一种可替代的来源，可用于治疗代谢性疾病和急性肝衰竭，作为肝移植的时间桥梁。这些细胞还可用于生物人工肝系统和药物毒性研究。
确定哪种冷保存溶液能更好地维持肝功能。
我们制备了4种由抗氧化剂、螯合剂、膜保护剂、抗凋亡剂以及亲环蛋白D抑制剂的不同组合制成的冷保存溶液。对于肝细胞分离，我们使用从未使用过的已故供体肝脏以及部分肝移植候选者的克里格勒 - 纳贾尔综合征患者的肝脏。培养和冷保存后，测量白蛋白水平和尿素生成量作为肝功能指标。
我们发现溶液1冷保存后白蛋白生成量显著下降。溶液1冷保存后24小时与对照组相比尿素生成量无显著差异。溶液2和溶液4冷保存后与对照组24小时相比白蛋白生成量无显著差异。溶液2和溶液4冷保存后与对照组24小时相比尿素生成量显著下降。总体而言，冷保存后白蛋白和尿素生成量显著下降。虽然冷保存后白蛋白和尿素生成量下降，但两种溶液的白蛋白生成结果与对照组无显著差异（p = 0.109和0.951）。
在我们的实验中，溶液2和溶液4对培养的人肝细胞的冷保存比其他冷保存溶液能更好地维持白蛋白生成功能；溶液1在4℃下对培养的人肝细胞尿素生成24小时更有效。为确定这些肝细胞是否适合移植，应进行进一步研究。
