Subcellular Fractionation of Human Intestinal Mucosa by Large-Scale Zonal Centrifugation

I. Characterization of Subcellular Organelles in the Distal Part of the Ileum

C. Tagesson Dept. of Clinical Chemistry; Dept. of Surgery, Unit for Gastroenterology; Dept. of Medical Microbiology; and Dept. of Pathology II; University of Linköping, Linköping, Sweden

T. Bolin Dept. of Clinical Chemistry; Dept. of Surgery, Unit for Gastroenterology; Dept. of Medical Microbiology; and Dept. of Pathology II; University of Linköping, Linköping, Sweden

R. Heuman Dept. of Clinical Chemistry; Dept. of Surgery, Unit for Gastroenterology; Dept. of Medical Microbiology; and Dept. of Pathology II; University of Linköping, Linköping, Sweden

K.-E. Magnusson Dept. of Clinical Chemistry; Dept. of Surgery, Unit for Gastroenterology; Dept. of Medical Microbiology; and Dept. of Pathology II; University of Linköping, Linköping, Sweden

K. Norrby Dept. of Clinical Chemistry; Dept. of Surgery, Unit for Gastroenterology; Dept. of Medical Microbiology; and Dept. of Pathology II; University of Linköping, Linköping, Sweden

R. Sjödahl Dept. of Clinical Chemistry; Dept. of Surgery, Unit for Gastroenterology; Dept. of Medical Microbiology; and Dept. of Pathology II; University of Linköping, Linköping, Sweden

Abstract

The subcellular organelles in human ileal mucosa have been characterized by largescale rate zonal and isopycnic density-gradient centrifugation. Mucosal scrapings were obtained from surgical specimens and the cells disrupted by extrusion under controlled pressure. The homogenized cells were then centrifuged to prepare a cell extract and the extract fractionated in a single-step procedure by zonal centrifugation on a continuous sucrose gradient. The subcellular organelles (brush borders, basallateral membranes, lysosomes, mitochondria, and endoplasmic reticulum) were located in the density gradient by assay of marker enzymes. Since a large number of analyses can be done on each fraction of interest, this fractionation technique may be used to study a variety of compounds and concepts involved in the subcellular physiology and pathology of human ileal mucosa.

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Subcellular Fractionation of Human Intestinal Mucosa by Large-Scale Zonal Centrifugation

I. Characterization of Subcellular Organelles in the Distal Part of the Ileum

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Subcellular Fractionation of Human Intestinal Mucosa by Large-Scale Zonal Centrifugation

I. Characterization of Subcellular Organelles in the Distal Part of the Ileum

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