The Growth Medium Affects the Viability of IPEC-J2 Animal Cell Line in the Presence of Probiotic Bacteria

Abstract

Background: The IPEC-J2 cell line is frequently used as an in vitro model to study the bioactivity of live probiotics. However, lactic acid bacteria (LB) acidify the medium, and the impact of pH and lactic acid accumulation on cell viability seem to be underestimated. Methods: IPEC-J2 viability was assessed by neutral red and flow cytometry in the presence of eight probiotics at concentrations between 106 and 109 bacteria/mL in maintenance and buffered media. Results: It was shown that a high inoculation level led to higher cytotoxic effects on IPEC-J2 cells after 22 h of incubation and that viability losses were more related to a combination of low pH and lactic acid than to the probiotics themselves. Furthermore, with LB at 106 and 107 bacteria/mL, the addition of phosphates to the media significantly reduced the drop in the pH and preserved the IPEC-J2 viability between 100% and 69%, compared to a highly variable viability between 100% and 17.5% in the unbuffered media. Conclusions: Under certain in vitro conditions, probiotics can lead to the deterioration of animal cells, and pH neutralization is an essential parameter in the cell–probiotic system in order to preserve cell viability and to better evaluate the bioactive properties of live probiotics.