Variation of the HES Concentration for the Cryopreservation of Keratinocytes in Suspensions and in Monolayers

Abstract

It has recently been shown that keratinocytes, both in suspension and in monolayers, can be successfully cryopreserved with hydroxyethyl starch (HES) (6, 9). HES is a nontoxic biodegradable macromolecule which is clinically approved as a plasma expander and which has already been used for the cryopreservation of red blood cells (10, 11). In this study we varied the HES concentration between 0 and 10 wt% in 2% steps for suspended cells and between 0, 4, 6, 8, and 10 wt% for monolayer cells in order to determine the effect on the survival rate and metabolic activity after cryopreservation. The experiments with the suspended cells were performed both with and without NCS. Cryopreserved keratinocytes can be transplanted onto patients for the treatment of deep dermal burns and leg ulcers. In this study, we achieved a survival rate of 80% for the suspended cells (10 wt% HES, 3°C/min) and a survival rate of even 88% when the cells were cryopreserved as a monolayer using the same parameters. The addition of NCS did not improve the results for the suspended cells significantly.