The Optimization of Short-Term Hepatocyte Preservation Before Transplantation

Abstract

BackgroundNo 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.MethodsHepatocytes 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.ResultsThe 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.ConclusionsTD is effective for short-term hepatocyte preservation. HSA is a useful substitute for FBS, and preserving in HM at low temperature is recommended.

Improvement of hepatocyte engraftment by co‐transplantation with pancreatic islets in hepatocyte transplantation

“…Hepatocytes were isolated from F344/NSLc rats by the two‐step collagenase perfusion as described previously (Fukuoka et al., 2017; Matsumura et al., 2019). First, Ca 2+ ‐free Hanks' balanced salt solution (HBSS, Sigma‐Aldrich) containing ethylene glycol tetra‐acetic acid was perfused through the portal vein at a rate of 14 ml/min for 5 min.…”

“…In spite of these advantages, the clinical application of HTx still faces many obstacles, including hepatocyte isolation, graft preservation (Fukuoka et al., 2017), graft quality evaluation (Matsumura et al., 2019), and hepatocyte engraftment (Domen, 2018; Weber, Groyer‐Picard, Franco, & Dagher, 2009). Among them, extremely poor engraftment of hepatocytes clearly remains the most severe issue.…”

A Simple and Useful Predictive Assay for Evaluating the Quality of Isolated Hepatocytes for Hepatocyte Transplantation

“…Although the results of PE and the DNA quantity were well-correlated with the HTx outcome, several days are required before both evaluations can be completed. Given that the isolated hepatocytes are fairly fragile 29 , these assays are not practical for the clinical setting. Alternatively, they may be effective for increasing the accuracy of quality assessment of isolated hepatocytes retrospectively in the research setting.…”

“…In the laboratory setting, studies of short-term drug metabolism and long-term cultured hepatocytes are commonly used to evaluate the quality of transplanted hepatocytes 26–28 . However, considering the fragility of isolated hepatocytes 29 , these assays require relatively too much time to assess the quality of grafts before HTx in the clinical settings; optimal methods must be established.…”

A linear shear model of cell viability loss during hepatocyte transplantation

“…The patterns determined the location of live cells in the microchannel and the intensity of shear stresses on the cells during flow in the microchannel. The cell death results indicate the need for improved practices related to cell isolation (Enosawa, 2017), transportation, and storage (Fukuoka et al, 2017) before injection for transplantation.…”