Trehalose cryoprotective
03 January 2007 - 22:37 Filed in:
PublicationsThe paper Cryopreservation of primary human hepatocytes – the benefit of trehalose as an additional cryoprotective agent by Ekaterina Katenz, Florian W.R. Vondran, Ruth Schwartlander, Gesine Pless, Xiaobing Gong, Xiandong Cheng, Peter Neuhaus and Igor M. Sauer is published in the January 2007 issue of Liver Transplantation.
Problems with the limited availability of human hepatocytes for cell transplantation may be overcome by efficient cryopreservation techniques and formation of appropriate cell banking. In the present study we investigated the effect of the disaccharide trehalose on the cryopreservation of human hepatocytes. For analysis, liver cells were frozen in culture medium containing 10% DMSO that was supplemented with varying concentrations of trehalose. During the post-thawing culture period, viability, plating efficiency, total protein, cell proliferation, enzyme leakage, albumin and urea formation as well as phase I and II metabolism were analyzed. In the pilot study, among the concentrations investigated, 0.2 M trehalose showed the best overall outcome. Compared to the use of DMSO alone, we found significant improvement in post-thaw cell viability (62.9 ± 13 vs. 46.9 ± 11 %, p < 0.01) and plating efficiency (41.5 ± 18 vs. 17.6 ± 13 %, p < 0.01) in the trehalose group. The use of trehalose as an additive for cryopreserving human hepatocytes resulted in a significantly increased total protein level in the attached cells, higher secretion of albumin and a lower AST level after thawing. The use of trehalose as cryoprotective agent significantly improves the outcome of human hepatocyte cryopreservation.
Tags: Cell Culture
