Fukuoka Kengo, Inagaki Akiko, Nakamura Yasuhiro, Matsumura Muneyuki, Yoshida Satoru, Imura Takehiro, Igarashi Yasuhiro, Miyagi Shigehito, Ohashi Kazuo, Enosawa Shin, Kamei Takashi, Unno Michiaki, Ohuchi Noriaki, Satomi Susumu, Goto Masafumi
Department of Surgery, Tohoku University Graduate School of Medicine, Sendai, Japan.
Division of Transplantation and Regenerative Medicine, Tohoku University School of Medicine, Sendai, Japan.
Transplant Direct. 2017 Jun 19;3(7):e176. doi: 10.1097/TXD.0000000000000687. eCollection 2017 Jul.
No optimal methods for short-term hepatocyte preservation have been established. We have recently developed a prominent oxygen-permeable bag (Tohoku Device [TD]) for pancreatic islet culture and transplantation. In this study, we investigated whether TD is also effective for hepatocyte preservation and tried to optimize other conditions.
Hepatocytes were preserved in the following conditions, and their outcomes were observed. First, the effectiveness of TD was investigated. Second, hepatocyte medium (HM) and organ preservation solutions with or without fetal bovine serum (FBS) were compared. Third, as supplementations, FBS and human serum albumin (HSA) were compared. Fourth, low, room and high temperature were compared. And finally, hepatocytes preserved in various conditions were transplanted into the subrenal capsule space of nonalbumin rats and engrafted areas were assessed.
The survival rate of hepatocytes preserved in TD tended to be higher and their viability and function were maintained significantly greater than those of non-TD group. Irrespective of FBS supplementation, the survival rate of HM group was significantly higher than those of organ preservation solution group while viabilities and plating efficiency were similar among them. Although survival rates of groups without FBS were extremely low, results of HSA supplemented group were not inferior to FBS supplemented group. Hepatocytes preserved at high temperature had the worst results. The engrafted area of TD group tended to be higher than those of other groups.
TD is effective for short-term hepatocyte preservation. HSA is a useful substitute for FBS, and preserving in HM at low temperature is recommended.
尚未建立短期肝细胞保存的最佳方法。我们最近开发了一种用于胰岛培养和移植的突出的透氧袋（东北装置[TD]）。在本研究中，我们调查了TD对肝细胞保存是否也有效，并试图优化其他条件。
在以下条件下保存肝细胞，并观察其结果。首先，研究TD的有效性。其次，比较有无胎牛血清（FBS）的肝细胞培养基（HM）和器官保存液。第三，作为补充剂，比较FBS和人血清白蛋白（HSA）。第四，比较低温、室温及高温。最后，将在各种条件下保存的肝细胞移植到无白蛋白大鼠的肾包膜下间隙，并评估植入面积。
保存在TD中的肝细胞存活率趋于更高，其活力和功能的维持明显优于非TD组。无论是否添加FBS，HM组的存活率均显著高于器官保存液组，而它们的活力和平板接种效率相似。虽然未添加FBS组肝细胞存活率极低，但添加HSA组的结果并不逊于添加FBS组。在高温下保存的肝细胞结果最差。TD组的植入面积趋于高于其他组。
TD对短期肝细胞保存有效。HSA是FBS的有用替代品，建议在低温下用HM保存。
