Studies of isolated and enriched rat antral mucosa gastrin cells |
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Authors: | Prof. Dr. W. G. Forssmann L. M. Lichtenberger V. Helmstaedter S. Ito |
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Affiliation: | (1) Departments of Anatomy, Harvard Medical School, Boston, Massachusetts, USA;(2) Anatomisches Institut III, Universität Heidelberg, Im Neuenheimer Feld 307, D-6900 Heidelberg, Germany |
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Abstract: | A technique has been developed to obtain viable, isolated and enriched populations of gastrin cells (G-cells) from the rat stomach. Restricted tissue samples from a small area of the pyloric antrum known to be particularly rich in G-cells, were sequentially digested with pronase followed by mechanical agitation, to remove the epithelial cells. This technique resulted in a significant enrichment of G-cells (3-4 fold) since the surface epithelial cells and upper portions of the glands were discarded before the initial G-cell fraction was collected. These cells in suspension were then isolated from each other by gentle pipetting in a DNase containing solution and designated the crude preparation (CP). The G-cells were then purified further by separating the cells according to size by velocity sedimentation. The greatest concentration of G-cells (15-25%) was found in the fraction containing cells with diameters of 10 to 12 micrometer. The effectiveness of the technique was evaluated by counting G-cells as identified by electron microscopy and immunofluorescence and assessing gastrin activity by radioimmunoassay. All three methods indicated that cell separation by gravity velocity sedimentation enriched the G-cell population 15-20 fold over their concentration in the CP. The combined techniques of selective pronase digestion followed by gravity velocity sedimentation resulted in an isolated cell preparation containing a 50-100 fold increase of G-cells over their normal distribution in the intact gastric mucosa. Since these isolated G-cells retain features indicating viability, their usefulness for in vitro studies is suggested. |
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Keywords: | Gastrin cells Entero-endocrine cells Rat Cell isolation Pylorus |
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