Effect of thermal variables on frozen human primary prostatic adenocarcinoma cells

Objectives. Recent advances in imaging technology and cryotechnology have rekindled interest in prostate cryosurgery. Cryosurgery, however, cannot be applied precisely without knowing how the thermal variables used during the procedure affect tissue destruction. The goal of this article is to provide quantitative values for the relationship between thermal variables during freezing and the destruction of human primary prostatic adenocarcinoma cells. Methods. Human primary prostatic adenocarcinoma cells were frozen with controlled thermal parameters, using a directional solidification apparatus. Cell viability was determined after thawing, using trypan blue and a two-dye fluorescent test and correlated to the thermal variables used during freezing. Results. Human primary prostatic adenocarcinoma cells are damaged by intracellular chemical damage when frozen with cooling rates lower than 5 degrees C/min and by intracellular ice formation when frozen with cooling rates higher than 25 degrees C/min. A double freeze/thaw cycle is required to ensure complete cell destruction at high subzero temperatures, which must be lower than -40 degrees C for the low cooling rates and lower than -19 degrees C for the higher cooling rate. Conclusions. Haphazard freezing does not necessarily destroy tissue during cryosurgery; however, quantitative data on the relation between thermal variables and frozen cell destruction can provide the means for performing cryosurgery more precisely and with greater control over the outcome of the procedure.