TY - JOUR
T1 - Isolation and purification of large quantities of fresh human Kupffer cells, which are cytotoxic against colon carcinoma
AU - Heuff, Gijsbert
AU - van de Loosdrecht, Arjan A.
AU - Betjes, Michiel G.
AU - Beelen, Robert H.J.
AU - Meijer, Sybren
PY - 1995/3
Y1 - 1995/3
N2 - A new rapid method is described for the isolation and purification of functional active human Kupffer cells without the need of in situ perfusion techniques. Liver wedge biopsies (3 to 5 g), obtained after laparotomy, were incubated with pronase under continuous pH registration. Human Kupffer cells were subsequently separated from other nonparenchymal cells by Nycodenz gradient centrifugation and purified by counterflow centrifugal elutriation. Kupffer cells, 1.7 +/- 0.4 x 10(6) per gram liver, were isolated with a purity of 95% +/- 3%. Cell-mediated cytotoxicity 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 increased to 73 +/- 17% (P < .05) after activation of Kupffer cells with interferon-gamma. In conclusion, a reliable and relatively simple method is provided to isolate and purify fresh human Kupffer cells in large yields, which show spontaneous as well as gamma-interferon-induced cytotoxicity against a human colon carcinoma cell line.
AB - A new rapid method is described for the isolation and purification of functional active human Kupffer cells without the need of in situ perfusion techniques. Liver wedge biopsies (3 to 5 g), obtained after laparotomy, were incubated with pronase under continuous pH registration. Human Kupffer cells were subsequently separated from other nonparenchymal cells by Nycodenz gradient centrifugation and purified by counterflow centrifugal elutriation. Kupffer cells, 1.7 +/- 0.4 x 10(6) per gram liver, were isolated with a purity of 95% +/- 3%. Cell-mediated cytotoxicity 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 increased to 73 +/- 17% (P < .05) after activation of Kupffer cells with interferon-gamma. In conclusion, a reliable and relatively simple method is provided to isolate and purify fresh human Kupffer cells in large yields, which show spontaneous as well as gamma-interferon-induced cytotoxicity against a human colon carcinoma cell line.
UR - https://aasldpubs.onlinelibrary.wiley.com/doi/pdf/10.1002/hep.1840210322
U2 - 10.1016/0270-9139(95)90527-8
DO - 10.1016/0270-9139(95)90527-8
M3 - Article
C2 - 7875672
SN - 0270-9139
VL - 21
SP - 740
EP - 745
JO - Hepatology
JF - Hepatology
IS - 3
ER -