Poloxamer 188 supplemented culture medium increases the vitality of Caco-2 cells after subcultivation and freeze/thaw cycles

Abstract

Cryostorage media containing 1% of the non-ionic surfactant Poloxamer 188 provided full recovery of mammalian cells (González Hernández, 2006), but its effects during thawing of cryostored cells and proteo­lytic subcultivation are still unknown. In this study, the proliferation and viability of pre-confluent Caco-2 monolayers cultivated in media supplemented with the non-ionic surfactant were investigated. The addition of 0.5% Poloxamer 188 increased proliferation of subcultivated cells 1.5 fold and that of thawed cells about twofold. According to microaspiration experiments the non-ionic surfactant increased the tension of the cell membrane most notably at concentrations ≤ 0.5% owing to adsorption and incorporation into the phospholipid bilayer. Thus, the performance of the cells appears to be improved. Since vitality of cells is a prerequisite for reproducibility and reliability of cell models for absorption studies at early stages of drug development, use of Poloxamer 188 supplemented cultivation media may help to refine cell culturing to further reduce animal trials in preclinical investigations.