Crofton R W, Diesselhoff-den Dulk M M, van Furth R
J Exp Med. 1978 Jul 1;148(1):1-17. doi: 10.1084/jem.148.1.1.
Enzymatic digestion with pronase and DNAase was used to isolate Kupffer cells from mouse liver. The characteristics of these cells were found to be similar to those of peritoneal macrophages, except that in the initial suspension the percentage of Kupffer cells with Fc receptors was low, C receptors were absent and the ingestion of opsenized bacteria was very poor, because of the effect of pronase on the cell membrane. After 24 h incubation in vitro all these characteristics return. The in vitro and 1 h-pulse [(3)H]thymidine labeling of the Kupffer cells is low (0.8 and 1 percent, respectively) indicating that in essence these cells do not divide. It was also shown that the small percentage of in vitro labeled Kupffer cells was recently derived from the circulation. After an intravenous injection of zymosan the in vitro labeling index of the Kupffer cells increased 16-fold, but it was proven that these dividing cells were immature mononuclear phagocytes very recently recruited from the bone marrow. The labeling of Kupffer cells aider one or four injections of [(3)H]thymidine reached a peak of 10.4 percent at 48 h or 24.1 percent at 60 h, respectively, indicating that these cells are derived from labeled monocytes. Further evidence for this conclusion was obtained by the absence of an increase of labeled Kupffer cells during treatment with hydrocortisone, which causes a monocytopenia during which no circulating monocytes are available to migrate to the tissues. Labeling studies in animals X-irradiated with hind-limb shielding gave a Kupffer cell labeling index of 5-10 percent of the normal values, which confirms their bone marrow origin. A quantitative study on the production of labeled monocytes in the bone marrow and their transit through the circulation showed that in the normal steady state at least 56.4 percent of the monocytes leaving the circulation become Kupffer cells. Considering the Kupffer cells as kinetically homogeneous this gives a mean turnover time of the total population of Kupffer cells of 21 days.
用链霉蛋白酶和脱氧核糖核酸酶进行酶消化，从小鼠肝脏中分离库普弗细胞。发现这些细胞的特性与腹腔巨噬细胞相似，只是在最初的悬浮液中，具有Fc受体的库普弗细胞百分比低，没有C受体，吞噬调理过的细菌能力很差，这是由于链霉蛋白酶对细胞膜的作用。体外培养24小时后，所有这些特性恢复。库普弗细胞的体外和1小时脉冲[³H]胸腺嘧啶核苷标记率很低（分别为0.8%和1%），表明这些细胞本质上不分裂。还表明，体外标记的库普弗细胞中一小部分是最近从循环中衍生而来的。静脉注射酵母聚糖后，库普弗细胞的体外标记指数增加了16倍，但已证明这些分裂细胞是最近从骨髓中招募的未成熟单核吞噬细胞。注射一次或四次[³H]胸腺嘧啶核苷后，库普弗细胞的标记率分别在48小时达到10.4%的峰值或在60小时达到24.1%的峰值，表明这些细胞源自标记的单核细胞。在用氢化可的松治疗期间，标记的库普弗细胞没有增加，这进一步证明了这一结论，因为氢化可的松会导致单核细胞减少，在此期间没有循环单核细胞可迁移到组织中。对后肢屏蔽X射线照射的动物进行的标记研究显示，库普弗细胞标记指数为正常值的5 - 10%，这证实了它们的骨髓来源。对骨髓中标记单核细胞的产生及其在循环中的转运进行的定量研究表明，在正常稳态下，至少56.4%离开循环的单核细胞变成库普弗细胞。将库普弗细胞视为动力学上同质的群体，这给出了库普弗细胞总数的平均更新时间为21天。
