ISOLATION AND PURIFICATION OF LARGE QUANTITIES OF FRESH HUMAN KUPFFERCELLS, WHICH ARE CYTOTOXIC AGAINST COLON-CARCINOMA

A new rapid method is described for the isolation and purification of functional active human Kupffer cells without the need of in situ perf usion techniques, Liver wedge biopsies (3 to 5 g), obtained after Lapa rotomy, were incubated with pronase under continuous pH registration. Human Kupffer cells were subsequently separated from other nonparenchy mal cells by Nycodenz gradient centrifugation and purified by counterf low centrifugal elutriation. Kupffer cells, 1.7 +/- 0.4 x 10(6) per gr am liver, were isolated with a purity of 95% +/- 3%. Cell-mediated cyt otoxicity of Kupffer cells was assayed against a human colon carcinoma cell Line (SW948). Kupffer cell cytotoxicity was 42% +/- 9% (mean +/- SD) at an effector-to-target cell ratio of 10 and significantly incre ased to 73 +/- 17% (P < .05) after activation of Kupffer cells with in terferon-gamma. In conclusion, a reliable and relatively. simple metho d is provided to isolate and purify fresh human Kupffer cells in Large yields, which show spontaneous as well as gamma-interferon-induced cy totoxicity against a human colon carcinoma cell line.