Use of a collagen membrane to enhance the survival of primary intestinal epithelial cells

FIORELLA DI CLAUDIO; MARÍA LUCÍA ORSINI DELGADO; GUILLERMO DOCENA; CECILIA MUGLIA; FERNANDO TREJO; PAOLA SMALDINI; RAÚL GRIGERA

JOURNAL OF CELLULAR PHYSIOLOGY

WILEY-LISS, DIV JOHN WILEY & SONS INC

Año: 2016

0021-9541

Intestinal epithelial cell culture is important for biological, functional and immunological studies.Since enterocytes have a short in vivo life span due to anoikis, we aimed to establish a novel andreproducible method to prolong the survival of mouse and human cells. Cells were isolatedfollowing a standard procedure, and cultured on ordered-cow´s collagen membranes. A prolongedcell life span was achieved; cells covered the complete surface of bio-membranes and showed aclassical enterocyte morphology with high expression of enzymes supporting the possibility ofcryopreservation. Apoptosis was dramatically reduced and cultured enterocytes expressedcytokeratin and LGR5 (low frequency). Cells exposed to LPS or flagellin showed the induction ofTLR4 and TLR5 expression and a functional phenotype upon exposure to the probioticBifidobacterium bifidum or the pathogenic Clostridium difficile. The secretion of the homeostatic(IL-25 and TSLP), inhibitory (IL-10 and TGF-β) or pro-inflammatory mediators (IL-1β and TNF)were induced. In conclusion, this novel protocol using cow´s collagen-ordered membrane providesa simple and reproducible method to maintain intestinal epithelial cells functional for cellmicroorganisminteraction studies and stem cell expansion.