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Enrichment of macrophages in cell suspensions of human intestinal mucosa by elutriation centrifugation.

作者信息

Beeken W, Mieremet-Ooms M, Ginsel L A, Leijh P C, Verspaget H

出版信息

J Immunol Methods. 1984 Oct 12;73(1):189-201. doi: 10.1016/0022-1759(84)90044-9.

Abstract

To obtain macrophage-rich cell suspensions from human intestinal mucosa, lamina propria specimens were dissociated by incubating in EDTA-collagenase-DNAase solutions and further purified by counter-current centrifugation. During enzymatic incubation macrophage dissociation was linear over the first 8-10 h, reaching a maximum concentration of 10% of total cells and then it plateaued. Counter-current centrifugation resulted in a 5-fold enrichment of macrophages to a mean of 50% with an average recovery rate of 84%. Yields exceeded 0.69 X 10(5) macrophages/g mucosa. Greater than 90% of these cells phagocytosed Staphylococcus aureus, and could be maintained in culture for up to 8 days. Electron microscopy showed satisfactory preservation of the ultrastructure of the macrophages, which also seemed functionally intact.

摘要

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