raffinose and Liver-Neoplasms

Primary human hepatocytes are clinically used for transplantation or in bioartificial liver devices for the treatment of patients with liver failure. We aimed to assess whether an organ preservation solution containing trehalose, namely ET-Kyoto solution (ETK), could improve human liver cell viability when used for cryopreservation in comparison to the University of Wisconsin solution (UW). Our study showed beneficial effects of ETK when used in combination with other cryoprotectants on the viability of thawed hepatocytes. Indeed, no significant difference was seen between the viability of freshly isolated cells and cryopreserved cells when cryopreserved with ETK combined with other cryoprotectants. In contrast, a significant decrease of viability was observed in cells cryopreserved with UW or ETK combined with dimethysulfoxide (DMSO) only, or with UW combined with other cryoprotectants, compared to freshly isolated cells. No significant difference was observed between the four different groups of cryopreserved hepatocytes with regards to cell recovery rate or cell attachment after thawing. However, a significant decrease in cell metabolic activity was found in cells cryopreserved with UW 10% DMSO compared to the other groups. In conclusion, our study confirms the beneficial effect of ETK for the cryopreservation of human hepatocytes in combination with other cryoprotective agents.

Topics: Adenosine; Adult; Aged; Allopurinol; Cell Culture Techniques; Cell Survival; Female; Gluconates; Glutathione; Hepatectomy; Hepatocytes; Humans; Hydroxyethyl Starch Derivatives; Insulin; Liver Neoplasms; Male; Middle Aged; Organ Preservation Solutions; Phosphates; Raffinose; Tissue Preservation; Trehalose

University of Wisconsin (UW) and histidine-tryptophan-ketoglutarate (HTK) solutions are the 2 most commonly used liver preservation solutions. The aim of this study was to compare cardiovascular stability, acid-base status, and potassium concentrations between patients who received grafts preserved in either UW or HTK solution in orthotopic liver transplantation (OLT).. In this retrospective study, 87 patients who underwent living donor OLT were divided into 2 groups: UW (n = 28) and HTK (n = 59). Group HTK was subdivided into group NF-HTK (n = 31; nonflushed before reperfusion) and group F-HTK (n = 28; flushed before reperfusion). We determined mean arterial pressure (MAP) and heart rate every minute for 5 minutes after reperfusion and the maximum change in these values and incidence of postreperfusion syndrome (PRS). Body temperature, cardiovascular and acid-base parameters, as well as potassium concentrations were compared at 5 minutes before and 5 and 30 minutes after reperfusion.. The maximum decreases in MAP within 5 minutes after reperfusion were significantly greater in both the NF-HTK and the F-HTK groups. The rate of PRS was significantly greater in the NF-HTK compared with the UW group. Flushing with HTK solution decreased the rate of PRS; there was no significant difference between the F-HTK and UW groups. All serial changes in body temperature, cardiovascular and acid-base parameters, as well as potassium concentrations were similar among the 3 groups.. The incidence of PRS was greater using HTK compared with UW solution during the reperfusion period. Therefore, careful hemodynamic management is advised when using HTK solution.

Topics: Acid-Base Equilibrium; Adenosine; Adult; Allopurinol; Blood Pressure; Carcinoma, Hepatocellular; Female; Glucose; Glutathione; Hemodynamics; Humans; Insulin; Liver Cirrhosis; Liver Neoplasms; Liver Transplantation; Living Donors; Male; Mannitol; Middle Aged; Organ Preservation Solutions; Portal Vein; Potassium; Potassium Chloride; Procaine; Raffinose; Reperfusion Injury; Retrospective Studies

Cell hydration changes critically affect liver metabolism and gene expression. In the course of gene expression studies using nylon cDNA-arrays we found that hyperosmolarity (405 mosmol/l) suppressed the betaine-homocysteine methyltransferase (Bhmt) mRNA expression in H4IIE rat hepatoma cells. This was confirmed by Northern blot and real-time quantitative RT-PCR analysis, which in addition unraveled a pronounced induction of Bhmt mRNA expression by hypoosmotic (205 mosmol/l) swelling. Osmotic regulation of Bhmt mRNA expression was largely paralleled at the levels of Bhmt protein and enzymatic activity. Like hyperosmotic NaCl, hyperosmotic raffinose but not hyperosmotic urea suppressed Bhmt mRNA expression, suggesting that cell shrinkage rather than increased ionic strength or hyperosmolarity per se is the trigger. Hypoosmolarity increased the expression of a reporter gene driven by the entire human BHMT promoter, whereas destabilization of BHMT mRNA was observed under hyperosmotic conditions. Osmosensitivity of Bhmt mRNA expression was impaired by inhibitors of tyrosine kinases and cyclic nucleotide-dependent kinases. The osmotic regulation of BHMT may be part of a cell volume-regulatory response and additionally lead to metabolic alterations that depend on the availability of betaine-derived methyl groups.

Topics: Animals; Betaine; Betaine-Homocysteine S-Methyltransferase; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Size; Cyclic Nucleotide-Regulated Protein Kinases; Gene Expression Regulation, Enzymologic; Liver Neoplasms; Osmolar Concentration; Osmosis; Promoter Regions, Genetic; Protein Kinase Inhibitors; Protein-Tyrosine Kinases; Raffinose; Rats; RNA, Messenger; Saline Solution, Hypertonic; Sarcosine; Signal Transduction; Time Factors; Transcription, Genetic; Transfection; Urea; Water-Electrolyte Balance

Topics: Adenosine; Allopurinol; Blood Loss, Surgical; Constriction; Embolism, Air; Follow-Up Studies; Glutathione; Hepatectomy; Hepatic Veins; Humans; Hypothermia, Induced; Insulin; Ischemia; Ligation; Liver Circulation; Liver Diseases; Liver Failure; Liver Neoplasms; Organ Preservation Solutions; Perfusion; Portal Vein; Raffinose; Survival Rate; Time Factors; Tissue Preservation; Vena Cava, Inferior

Topics: Adenoma, Bile Duct; Adenosine; Adult; Allopurinol; Bile Duct Neoplasms; Carcinoma, Hepatocellular; Colonic Neoplasms; Duodenal Neoplasms; Duodenum; Female; Glutathione; Humans; Insulin; Intestine, Small; Liver Neoplasms; Liver Transplantation; Male; Middle Aged; Organ Preservation; Organ Preservation Solutions; Pancreas Transplantation; Pancreatic Neoplasms; Raffinose; Solutions