In vitro biocompatibility performance of Physioneal. toneal dialysis (PD) has been a successful and effective form of chronic renal replacement therapy since its introduction over 20 years ago. Despite its overall success, there is a growing body of evidence that suggests shortcomings in the preservation of membrane integrity. This has led to the development of several second-generation PD solutions that demonstrate improved biocompatibility. Physioneal, a neutral pH, bicarbonate/lactate-buffered solution, was one of the first of these new PD solutions to become commercially available. This review will focus on one of the first preclinical stages in the development of Physioneal: studies on in vitro biocompatibility testing. Studies in leukocyte, mesothelial cell, and fibroblast populations demonstrated significantly improved biocompatibility of neutral pH, bicarbonate/lactate-based solutions compared to conventional solutions. The solutions contributed to improved leukocyte viability and response to bacterial infection (e.g., phagocytosis, superoxide radical generation, and endotoxin-stimulated cytokine release). Studies on peritoneal mesothelial cells demonstrate improved cell viability, proliferation, and response to proinflammatory stimuli, and a reduced potential for angiogenesis and peritoneal fibrosis, all suggesting a better preservation of membrane structure and function. The bicarbonate/lactate-based solutions demonstrated decreased cytotoxicity and preserved cell growth in fibroblast cultures as well. In vitro biocompatibility testing has clearly demonstrated that neutral pH, bicarbonate/lactate-buffered Physioneal solutions are superior to conventional solutions in preserving cell viability and function in cell populations that contribute to peritoneal homeostasis. This positive assessment now provides a foundation and rationale for moving forward with the next stages in preclinical testing: in vivo animal models and human ex vivo studies.