Background: Intraoperative tumour spillage can be concerning during cancer excisions, given it can lead to tumour-cell re-implantation and local recurrence. Examples include bladder tumour recurrences post-transurethral resection, or peritoneal spillage during laparotomy/laparoscopy for bowel and ovarian cancers. One approach to reducing implantation is mechanical wash out of free-floating tumour cells. Irrigation with water may have additional effectiveness compared to iso-osmotic irrigants (e.g. saline) by causing osmotic cytolysis, but this is not well-characterised. This in vitro study aimed to ascertain the time-course of osmotic effects of water on various cancer cell lines to provide guidance for clinical usage. Methods: Assays were conducted on six cancer cell lines (bladder [HT1197, HT1376], colon [KM12, LIM2405], kidney [SKRC52], and ovarian [COV434]). Cells were exposed to water or 0.9% saline and cell counts were performed using a haemocytometer at 10, 20, 40, 60, 120 and 180 min. Cell viability was determined using Trypan Blue exclusion.Results: In all cell lines, exposure to water led to 100% cell lysis within a median time of 40 min (range 10-180 min), while exposure to saline led to a gradual decline in cell viability (median 50.2%, range 6.7%-100.0%) over 3 h, and did not result in complete cell lysis. An increase in osmotic gradient equivalent to a concentration of 5% NaCl was sufficient to impede the effects of water-mediated cell lysis. Conclusion: Our studies suggest that water has a rapid osmolytic effect on cancer cells. The required exposure time to reach 0% cell viability varied between individual cell lines.