IngentaConnect Integrity and metabolism of human ileal mucosa in vitro in the Us...

Authors: SÖDERHOLM¹; HEDMAN¹; ARTURSSON²; FRANZE´N³; LARSSON⁴; PANTZAR⁵; PERMERT⁴; OLAISON¹

Source: Acta Physiologica, Volume 162, Number 1, January 1998 , pp. 47-56(10)

Abstract:

The Ussing chamber is increasingly being used for in vitro studies of human intestinal mucosa, but little attention has been paid to the viability of specimens over time. Ninety-one mucosal specimens from the ileum in 19 patients operated on for colonic cancer were studied in regard to intestinal barrier function, metabolism, electrophysiology and histology during 360 min of incubation in Ussing chambers. Steady-state permeability to ⁵¹Cr-EDTA was maintained for 120 min. Mucosal ATP and lactate levels were stable for 180 min and transmucosal glucose flux for 240 min. Lactate dehydrogenase leakage was limited within 120 min. Transepithelial potential difference was 9.0 ± 3.0 mV at the start, and declined slowly throughout 360 min. Light microscopy revealed epithelial lifting from the basal lamina at 90 min. Transmission electron microscopy demonstrated preserved ultrastructure for 120 min. Specimens with a transepithelial potential difference below 6 mV at the start were associated with increased ⁵¹Cr-EDTA permeability and lactate dehydrogenase leakage and more pronounced light microscopy changes. All studied parameters pointed to preserved viability if experiments were kept within a period of 90 min after equilibration. The few specimens with early viability derangement were identified by a transepithelial potential difference below 6 mV at the start. The Ussing chamber provides a tool for in vitro studies of human intestinal epithelium, including permeability. To minimize viability problems, experiments should be limited in time and monitored by measurements of transepithelial potential difference.

Keywords: diffusion chamber; histology; human; ileum; intestinal absorption; intestinal mucosa; in vitro; tissue survival

Document Type: Original article

DOI: 10.1046/j.1365-201X.1998.0248f.x

Affiliations: 1: Department of Surgery and Clinical Research Center, University Hospital, Linköping, Sweden 2: Department of Pharmaceutics, University of Uppsala, Sweden 3: Departments of Pathology, University Hospital, Linköping and Ryhov Hospital, Jönköping, Sweden 4: Department of Surgery, Huddinge University Hospital, Karolinska Institute, Stockholm, Sweden 5: Department of Animal Physiology, University of Lund, Sweden

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Integrity and metabolism of human ileal mucosa in vitro in the Ussing chamber