Fisker S, Kudahl K, Sonne O
Institute of Physiology, University of Aarhus, Denmark.
J Immunol Methods. 1990 Oct 4;133(1):31-8. doi: 10.1016/0022-1759(90)90315-m.
Centrifugation of rat leucocytes from thioglycollate-induced inflammatory peritoneal exudate on a discontinuous gradient of Nycodenz with a density of 1106 g/l and an osmolarity of 400 mosmol/l separated the polymorphonuclear from the mononuclear leucocytes. The cells on the interphase between the buffer and the gradient medium contained 96% mononuclear leucocytes with a recovery of greater than 60%, and the bottom fraction consisted of 98% polymorphonuclear leucocytes with a yield of 91%, when an exudate isolated 20 h after the injection of thioglycollate was fractionated. Leucocytes isolated from non-inflamed rat peritoneum could be enriched in the fraction of nonspecific esterase-positive cells from 86% to 96% with a recovery of 82% on a gradient with a density of 1091 g/l and an osmolarity of 325 mosmol/l. The viability of the isolated cells was greater than 95% (trypan blue exclusion test), and there was no measurable reduction in the fraction of phagocytosing cells (latex and opsonized zymosan) after exposure to the hypertonic gradient material.
用密度为1106 g/l、渗透压为400 mosmol/l的不连续Nycodenz梯度对硫代乙醇酸盐诱导的炎性腹膜渗出液中的大鼠白细胞进行离心，可将多形核白细胞与单核白细胞分离。当对注射硫代乙醇酸盐20小时后分离的渗出液进行分级分离时，缓冲液与梯度介质界面处的细胞含有96%的单核白细胞，回收率大于60%，底部部分由98%的多形核白细胞组成，产率为91%。从非炎性大鼠腹膜分离的白细胞在密度为1091 g/l、渗透压为325 mosmol/l的梯度上，非特异性酯酶阳性细胞部分可从86%富集到96%，回收率为82%。分离细胞的活力大于95%（台盼蓝排斥试验），暴露于高渗梯度材料后，吞噬细胞（乳胶和调理酵母聚糖）的比例没有可测量的降低。
