Maynard J R, Fintel D J, Pitlick F A, Nemerson Y
Lab Invest. 1976 Dec;35(6):542-9.
Tissue factor content of WISH amnion cells in spinner culture increases 3- to 10-fold within 12 hours after subculture, then declines to a basal level within 30 to 50 hours. Maximal development of activity requires fresh serum and fresh medium. When added at the time of subculture, actinomycin D and cycloheximide completely inhibit development of coagulant activity; when added several hours after transfer, these inhibitors suppress the development but do not affect the disappearance of activity. Of the oxidative phosphorylation inhibitors tested, dinitrophenol had no effect whereas carbonyl cyanide m-chlorophenylhydrazone inhibited the activity increase but did not alter the decline. The kinetics of development and decay are similar over a pH range of 6.7 to 7.6 and with fetal calf serum concentration between 5 and 30 per cent. At pH 6.7 or in 30 per cent fetal calf serum, cell division did not occur. 3H-leucine and 35SO4= incorporation into the cell surface coat did not change appreciably during the burst of coagulant activity nor did the levels of naphthylamidase or alkaline phosphatase; 3H-thymidine incorporation reached a peak within 2 hours of the tissue factor maximum.
转瓶培养的WISH羊膜细胞的组织因子含量在传代培养后12小时内增加3至10倍,然后在30至50小时内降至基础水平。活性的最大发展需要新鲜血清和新鲜培养基。在传代培养时加入放线菌素D和环己酰亚胺可完全抑制凝血活性的发展;在转移后数小时加入,这些抑制剂会抑制其发展,但不影响活性的消失。在所测试的氧化磷酸化抑制剂中,二硝基苯酚没有作用,而羰基氰化物间氯苯腙抑制活性增加但不改变其下降。在pH值为6.7至7.6以及胎牛血清浓度在5%至30%之间时,发展和衰减的动力学相似。在pH值为6.7或30%胎牛血清中,细胞不发生分裂。在凝血活性爆发期间,3H-亮氨酸和35SO4=掺入细胞表面被膜的情况没有明显变化,萘基酰胺酶或碱性磷酸酶的水平也没有变化;3H-胸腺嘧啶核苷掺入在组织因子达到最大值后2小时内达到峰值。