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作者按照Tragager和Jensen的方法,用含10%人血清的RPMI 1640-HEPES(称为RPS培基)在100mm的陪氏培养皿中培养恶性疟原虫。为收集大量的裂殖子,首先进行同步培养。最初,用山梨糖醇处理培养物,然后,在明胶中漂浮反复地分离环状体和滋养体。用这种方法(一般是5次)每40—42小时挑选一次感染滋养体的红细胞,直到3—4小时内原虫同步发育。在释放裂殖子预测时间的八小时前,吸出培养基,并用含下列任一基质:(a)[~(35)S]蛋氨酸,10μci/ml的培养基;(b)[~3H]亮氨酸,10μci/ml 60ci/mmol;(c)[~3H]葡萄糖胺,40μci/ml;(d)[~3H]甘露糖,25μci/ml的10ml RPS培养基进行补充。
The authors cultured P. falciparum in 100 mm Petri dishes with RPMI 1640-HEPES (referred to as RPS medium) containing 10% human serum according to the method of Tragager and Jensen. To collect a large number of merozoites, the first synchronous culture. Initially, the cultures were treated with sorbitol and then the rings and trophozoites were repeatedly separated floating in gelatin. In this way (usually 5 times) every 40-42 hours to pick trophozoites red blood cells until protozoan synchronized development within 3-4 hours. Eight hours prior to the predicted time for merozoite aspiration, medium was aspirated and used with either of the following: (a) [~ (35) S] methionine, 10μci / ml medium; (C) [~ 3H] glucosamine, 40μci / ml; (d) [~ 3H] mannose, 25μci / ml in 10ml RPS medium for supplementation.