Organotypical Tissue Cultures from Fetal and Neonatal Murine Colon

Abstract

The complex functions of the gastrointestinal tract rely on the coordinated interplay of several cell and tissue types involving epithelium, connective tissue, smooth muscles as well as cells of the immune and nervous system. It is therefore obvious, that these functions can hardly be investigated sufficiently using cell lines or two-dimensional cell cultures.Here, we describe an easy to produce three-dimensional organotypical explants culture from fetal and neonatal murine colon. This model is suitable for in vitro testing of intestinal function or the evaluation of developmental or pathological processes.

Claudin expression in the rat endolymphatic duct and sac - first insights into regulation of the paracellular barrier by vasopressin

“…To test the regulatory effect of AVP on tight –junction proteins in the ED and ES epithelium, the V2R specific agonist 1-Desamino-8-D-Arginin-Vasopressin (dDAVP; Bachem, Bubendorf, Switzerland) in the range of physiological concentrations of 10 −6  M, 10 −8  M and 10 −10  M and of the V2R antagonist H-9400 (concentrations: 10 −6  M μM, 10 −8  M and 10 −10  M; Bachem, Bubendorf, Switzerland) were added to the DMEM/F12 medium without HEPES supplemented with 25% horse serum, 10 IU/ml penicillin/streptomycin, 2 mM L-glutamine, 0,9 mg/ml sodium bicarbonate (ThermoFisher Scientific)114, respectively. Incubation of ES-specimens was performed for 30 min, 1 h, and 3 h. The range of physiological concentration values115 and time periods were selected from former studies dealing with AVP stimulation experiments in the ES and the kidney100116117.…”