An amino acidic adjuvant to augment cryoinjury of MCF-7 breast cancer cells

Abstract

One of the major challenges in cryosurgery is to minimize incomplete cryodestruction near the edge of the iceball. In the present study, the feasibility and effectiveness of an amino acidic adjuvant, glycine was investigated to enhance the cryodestruction of MCF-7 human breast cancer cell at mild freezing/thawing conditions via eutectic solidification. The effects of glycine addition on the phase change characteristics of NaCl–water binary mixture were investigated with a differential scanning calorimeter and cryo-macro/microscope. The results confirmed that a NaCl–glycine–water mixture has two distinct eutectic phase change events – binary eutectic solidification of water–glycine, and ternary eutectic solidification of NaCl–glycine–water. In addition, its effects on the cryoinjury of MCF-7 cells were investigated by assessing the post-thaw cellular viability after a single freezing/thawing cycle with various eutectic solidification conditions due to different glycine concentrations, end temperatures and hold times. The viability of MCF-7 cells in isotonic saline supplemented with 10% or 20% glycine without freezing/thawing remained higher than 90% ( n = 9), indicating no apparent toxicity was induced by the addition of glycine. With 10% glycine supplement, the viability of the cells frozen to −8.5 °C decreased from 85.9 ± 1.8% to 38.5 ± 1.0% on the occurrence of binary eutectic solidification of glycine–water ( n = 3 for each group). With 20% glycine supplement, the viability of the cells frozen to −8.5 °C showed similar trends to those with 10% supplement. However, as the end temperature was lowered to −15 °C, the viability drastically decreased from 62.5 ± 2.0% to 3.6 ± 0.7% ( n = 3 for each group). The influences of eutectic kinetics such as nucleation temperature, hold time and method were less significant. These results imply that the binary eutectic solidification of water–glycine can augment the cryoinjury of MCF-7 cells, and the extent of the eutectic solidification is significant.