Furthermore, hepatocytes isolated from liver samples are produced acutely when a tissue is available and often in larger amounts than immediately needed. In many situations, cryopreservation is desired to ship hepatocytes between laboratories and hospitals. Hepatocytes can be successfully isolated from resected livers and from livers not suitable for transplantation. Hepatocytes are also needed for drug metabolizing enzyme induction studies in vitro. Hepatocyte transplantation may function as a bridge to liver transplantation when donors are not available, especially in hepatic emergencies such as acute liver failure. Hepatocyte transplantation is gaining more attention as an alternative today. Lack of donor organs, among others, is still a major obstacle. Liver transplantation is still the golden standard treatment for acute liver failure and end-stage liver disease.

Liver disease is a major health problem worldwide. Estimating CYP activities did not seem to be a relevant way to compare hepatocyte function between different groups due to high normal variability between different liver samples.Д¬ONCLUSION: The cryoprotectant combination may be a better alternative to the standard DMSO-UW protocol in primary human hepatocyte cryopreservation. Thus, it may be a better alternative to the standard DMSO-UW protocol. Cryopreservation of human hepatocytes using the cryoprotectant combination was a simple and xeno-free procedure yielding better hepatocyte viability. However, due to high within-group variability in the activities of the major CYPs, any statistical between-group differences were precluded. There was a correlation between viability of fresh hepatocytes and the difference in cell viability between CB and DMSO protocols ( r 2 = 0.69) using the TB method. There was no significant difference in viability estimation between both the trypan blue (TB) and the Live-Dead Assay methods. RESULTS: The new cryoprotectant combination preserved hepatocyte viability significantly better than the standard DMSO-UW protocol ( P < 0.01). The function of the hepatocytes was evaluated before and after cryopreservation by assessing enzymatic activity of 6 major cytochrome P450 isoforms (CYPs): CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4 and CYP3A7. The viability of the hepatocytes was assessed by the trypan blue exclusion method as well as a calcein-esterase based live-dead assay before and after cryopreservation.

Using the polystyrene box freezing, we compared two xeno-free freezing solutions for freezing of primary human hepatocytes: a new medium (STEM-CELLBANKER, CB), which contains dimethylsulphoxide (DMSO) and anhydrous dextrose, both permeating and non-permeating cryoprotectants, and the frequently used DMSO - University of Wisconsin (DMSO-UW) medium. We first compared controlled rate freezing to polystyrene box freezing and did not find any significant change between the groups. Despite several hepatocyte cryopreservation protocols being available, improvements are urgently needed. METHODS: The demand for cryopreserved hepatocytes is increasing for both clinical and research purposes.

AIM: To optimize a xeno-free cryopreservation protocol for primary human hepatocytes.

State vs. dmso jaikoz wilson