Abstract

Establishment and Characterization of a Newly Generated Cell Line based on Primary Small Intestinal Tissue

Christina Fey, PhD, Chair of Tissue Engineering and Regenerative Medicine

Standardized small intestinal in vitro models matching the human physiology are needed, to investigate uptake, metabolism and excretion of pharmaceutical substances or for infection studies with human obligate pathogens. Current models like the Caco-2 tumor cell line model, mouse in vivo models or human primary in vitro models either lack in standardization or in cellular diversity to recapitulate the small intestine properly. Therefore, primary small intestinal organoids from mice and human were immortalized by lentiviral transfection in collaboration with the company InSCREENeX. These cells revealed stable cell growth over several months, a characteristic epithelial cell morphology and depicted essential functional structures of an intact epithelium such as microvilli and tight-junctions. A mixed cell population of gut-specific epithelial cell types were detected on protein as well as gene expression level. Further physiological relevant properties were indicated by an increasing barrier integrity over 21 day’s in vitro culture and functional transporter activity. These cell clones also have the potential to grow as organoids in matrigel as known for primary cells. Data generated by these models will close a gap between existing in vivo and in vitro models, which might help to improve the interpretation of in vivo data from mice, in particular.