ENRICHMENT OF MACROPHAGES IN CELL-SUSPENSIONS OF HUMAN INTESTINAL-MUCOSA BY ELUTRIATION CENTRIFUGATION

被引:17
作者
BEEKEN, W
MIEREMETOOMS, M
GINSEL, LA
LEIJH, PCJ
VERSPAGET, H
机构
[1] LEIDEN STATE UNIV HOSP, DEPT INFECT DIS, 2333 AA LEIDEN, NETHERLANDS
[2] LEIDEN STATE UNIV HOSP, DEPT ELECTRON MICROSCOPY, 2333 AA LEIDEN, NETHERLANDS
[3] LEIDEN STATE UNIV HOSP, DEPT GASTROENTEROL, 2333 AA LEIDEN, NETHERLANDS
关键词
D O I
10.1016/0022-1759(84)90044-9
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
To obtain macrophage-rich cell suspensions from human intestinal mucosa, lamina propria specimens were dissociated by incubating in EDTA-collagenase-DNase solutions and further by counter-current centrifugation. During enzymatic incubation macrophage dissociation was linear over the 1st 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 .times. 105 macrophages/g mucosa. Greater than 90% of these cells phagocytosed Staphylococcus aureus and could be maintained in culture for up to 8 days. EM showed satisfactory preservation of the ultrastructure of the macrophages, which also seemed functionally intact.
引用
收藏
页码:189 / 201
页数:13
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