Evaluation of Various Cryoablation Protocols in Multiple Cell 1ines using Quantitative Biomarkers of Bioluminescence Imaging: An <i>in vitro</i> Study

Abstract

Background & Aims: To investigate whether the optimal treatment protocols of cryoablation differ among various cell lines utilizing bioluminescence imaging (BLI). Methods: Eight luciferase-expressing cancer cell lines originated from various organs were used in this study, including lung, renal, liver, breast, colon, prostate, and skin. These cell lines were uniformly seeded in 96-well plates. The cell lines were then divided into four groups: Group 0 (G0), as a control group, and three treatment groups (G1-G3). G1 -G3 were subjected to cryoablation at temperatures of -45℃, -60℃, and -80℃, respectively, with varying treatment cycles (single, double, or triple cycle, respectively). Each freeze-thaw cycle consisted of ten minutes of freezing followed by five minutes of thawing. Finally, the average radiance of the cells was assessed using BLI.Results: Cryoablation at "-60℃ and -80℃" showed a significant decrease in average radiance, even after a single cycle of treatment in all cell lines. The average radiance of cells treated with -45℃ was substantially lower than that of untreated cell lines except melanoma. Conclusion: We found that -45℃ successfully eradicated all cancer cells across multiple cycles except melanoma. The most effective cryoablation protocol for all cell lines was observed at temperatures of -60℃ or lower.