The effect of n-acetylcysteine on hepatic histomorphology during hypothermic preservation

Risso, Patricia Sayuri; Koike, Márcia Kiyomi; Abrahão, Marcos de Souza; Ferreira, Natalie Chaves; Montero, Edna Frasson de Souza

doi:10.1590/s0102-86502014001700006

Cited by 4 publications

(2 citation statements)

References 17 publications

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“…The pre-incubation of hepatocytes in a medium supplemented with sugars, insulin, reduced glutathione, and NAC prior to cryopreservation improves cell recovery after thawing [ 40 ]. Recent studies have investigated the effects of NAC in the pre-incubation stages of hepatocyte cryopreservation [ 40 ]; however, only a few studies have examined its role in the hypothermic preservation of hepatocytes and their cold storage [ 41 ].…”

Section: Discussionmentioning

confidence: 99%

Establishment of a Serum-Free Hepatocyte Cryopreservation Process for the Development of an “Off-the-Shelf” Bioartificial Liver System

et al. 2022

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To use hepatocytes immediately when necessary for hepatocyte transplantation and bioartificial liver (BAL) systems, a serum-free cryopreservation protocol ensuring the high survival of hepatocytes and maintenance of their functions should be developed. We established a serum-free protocol for the cryopreservation of primary hepatocytes, hepatocyte spheroids, and hepatocyte spheroid beads in liquid nitrogen. The serum-free cryopreservation solutions showed a significantly higher performance in maintaining enhanced viability and ammonia removal, urea secretion, and the albumin synthesis of hepatocyte spheroids and spheroid beads. The serum-free thawing medium, containing human serum albumin (HSA) and N-acetylcysteine (NAC), was compared with a fetal bovine serum-containing thawing medium for the development of a serum-free thawing medium. Our results show that hepatocyte spheroids and spheroid beads thawed using a serum-free thawing medium containing HSA and NAC exhibited increased hepatocyte viability, ammonia removal, urea secretion, and albumin synthesis compared to those thawed using the serum-containing medium. Finally, we evaluated the liver functions of the cryopreserved BAL system-applied serum-free cryopreservation process compared to the fresh BAL system. The ammonia removal efficiency of the cryopreserved hepatocyte spheroids BAL was lower than or similar to that of the fresh BAL system. Additionally, the urea concentrations in the media of all three BAL systems were not significantly different during BAL system operation. This cryopreserved spheroid-based BAL system using a serum-free process will be a good candidate for the treatment of patients.

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Section: Discussionmentioning

confidence: 99%

Establishment of a Serum-Free Hepatocyte Cryopreservation Process for the Development of an “Off-the-Shelf” Bioartificial Liver System

et al. 2022

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“…This evidence was collected by a pathologist in a blinded manner. Four fields per section, totaling 5000 mm 2 , were analyzed under 100x magnification using a computerized image system (Axiovision release 4.7/05-2008, Carl Zeiss) coupled to an optical microscope (Axios-kop40) 16 . The ratio of dead/injured hepatocytes to the total number of hepatocytes was calculated; cell injury/death was indicated by the presence of swelling and vacuolar degeneration.…”

Section: Histological Evaluation Of Cell Damagementioning

confidence: 99%

Phenothiazines Enhance the Hypothermic Preservation of Liver Grafts: A Pilot in Vitro Study

Yang

Stone

et al. 2019

Cell Transplant

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In vitro liver conservation is an issue of ongoing critical importance in graft transplantation. In this study, we investigated the possibility of augmenting the standard pre-transplant liver conservation protocol (University of Wisconsin (UW) cold solution) with the phenothiazines chlorpromazine and promethazine. Livers from male Sprague-Dawley rats were preserved either in UW solution alone, or in UW solution plus either 2.4, 3.6, or 4.8 mg chlorpromazine and promethazine (CþP, 1:1). The extent of liver injury following preservation was determined by alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, the ratio of AST/ALT, morphological changes as assessed by hematoxylin-eosin staining, apoptotic cell death as determined by ELISA, and by expression of the apoptotic regulatory proteins BAX and Bcl-2. Levels of glucose (GLU) and lactate dehydrogenase (LDH) in the preservation liquid were determined at 3, 12, and 24 h after incubation to assess glucose metabolism. Oxidative stress was assessed by levels of superoxide dismutase (SOD), reactive oxygen species (ROS), and malondialdehyde (MDA), and inflammatory cytokine expression was evaluated with Western blotting. CþP augmentation induced significant reductions in ALT and AST activities; the AST/ALT ratio; as well as in cellular swelling, vacuolar degeneration, apoptosis, and BAX expression. These changes were associated with lowered levels of GLU and LDH; decreased expression of SOD, MDA, ROS, TNF-a, and IL-1b; and increased expression of Bcl-2. We conclude that CþP augments hypothermic preservation of liver tissue by protecting hepatocytes from ischemia-induced oxidative stress and metabolic dysfunction. This result provides a basis for improvement of the current preservation strategy, and thus for the development of a more effective graft conservation method.

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Apoptosis

Bennuri¹,

Rose²,

Frye³

2018

The Therapeutic Use of N-Acetylcysteine (NAC) in Medicine

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The effect of n-acetylcysteine on hepatic histomorphology during hypothermic preservation

Cited by 4 publications

References 17 publications

Establishment of a Serum-Free Hepatocyte Cryopreservation Process for the Development of an “Off-the-Shelf” Bioartificial Liver System

Establishment of a Serum-Free Hepatocyte Cryopreservation Process for the Development of an “Off-the-Shelf” Bioartificial Liver System

Phenothiazines Enhance the Hypothermic Preservation of Liver Grafts: A Pilot in Vitro Study

Apoptosis

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