Real time analysis of the interaction of isolated intestinal microbiota and supernatants from different human population groups with a colon adenocarcinoma cell line

Abstract

[Backgrounds] The gut microbiota constitutes a complex ecosystem that changes through the lifespan. Our understanding on the functional characterisitcs of the microbiota is mainly based on in vitro models that focus in the analysis of the interaction between the host and specific bacterial strains. Therefore, the development of simple and economically affordable study models, which take into account the complexity and functionality of the intestinal microbiota is required. [Objectives] The ability of an in vitro model, based on the real-time monitoring of the HT29 colon adenocarcinoma cell line, was evaluated to asses its ability to detect differences in the functionality of faecal supernatants (FS) and isolated faecal microbiotas (M) from different human population groups: obese (n = 9), healthy normal weight adults (n = 7) and ulcerative colitis patients (n = 5). A group of infants (n = 20), divided in full-term infants (2 days of age, n = 10) and preterm infants of the same age (n = 10) were also studied. [Methods] The composition of the M was characterized by 16S rRNA gene sequencing. The quantification of short chain fatty acids was also determined by gas chromatography in the FS. Moreover, the effect of M and FS was monitored in real time, using RT-Cell Analyzer technology on the proliferation and confluence phases of the HT29 cell line. [Conclusions] The microbiota-host interaction in vitro model developed allowed to detect differences in the functionality of the microbiotas of different population groups. This in vitro model shows potentiality for the characterization of the functionality of complex microbiotas and the resulting effect of modulating them by probiotics, prebiotics or other functional components of diet.