ENRICHMENT OF MOUSE SPLENIC NATURAL-KILLER CELLS USING DISCONTINUOUS POLYVINYLPYRROLIDONE SILICA (PERCOLL) GRADIENTS

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  • 1 January 1984
    • journal article
    • research article
    • Vol. 53  (4) , 721-729
Abstract
A simple and rapid method is reported for enriching murine spleen cells with natural killer (NK) function as assessed by short-term cytolysis assay of 51Cr-labeled YAC-1 lymphoma target cells. A reproducible procedure for enriching mouse splenic NK cells was developed using a 4- or 5-step discontinuous Percoll gradient in the density range of 1.062 g/ml (top) to 1.092 g/ml (bottom) and osmolarity (310-340 mOsm/kg) nearer to that of mouse blood and tissue. A 4- to 25-fold (usually .apprx. 9-fold) increase in NK cell activity, consisting of 50-100% of the recovered lytic unit activity, is found in which are the cells forming band 3, .apprx. 10% of the recovered cell number. This cell population with enriched NK cell activity has a characteristic density .ltoreq. 1.077 g/ml, but > 1.070 g/ml when centrifuged under appropriate conditions. Similar enrichment was obtained with a 4-step gradient at a uniform osmolarity of 320 mOsm/kg throughout. Although the lymphocytes in band 3 show relatively little heterogeneity in appearance, only a minor population of the cells contain granules.