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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Continuous axenic cultivation of Pneumocystis carinii
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Continuous axenic cultivation of Pneumocystis carinii

机译：卡氏肺孢子虫的连续性不育培养

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摘要

Continuous axenic culture of Pneumocystis carinii has been achieved. A culture vessel is used that allows for frequent medium exchange without disturbance of organisms that grow attached to a collagen-coated porous mem- brane. The growth medium is based on Minimal Essential Medium with Earle's salt supplemented with S-adenosyl-L-methionine, putrescine, ferric pyrophosphate, N-acetyl glu-cosamine, putrescine, p-aminobenzoic acid, L-cysteine and L-glutamine, and horse serum. Incubation is in room air at 31 degrees C. The pH of the medium begins at 8.8 and rises to approximately equals 9 as the cells grow. Doubling times calculated from growth curves obtained from cultures inoculated at moderate densities ranged from 35 to 65 hours. With a low-density inoculum, the doubling time is reduced to l9 hours. The morphology of cultured organisms in stained smears and in transmission electron micrographs is that of P carinii, and P carinii- specific mAbs label the cultured material. Cultured organisms are infective for immunosuppressed rats and can be stored frozen and used to reinitiate culture.
机译：已经实现了卡氏肺孢子虫的连续性焦虑培养。使用的培养容器可进行频繁的培养基交换，而不会干扰附着在胶原蛋白涂层的多孔膜上生长的生物。生长培养基基于添加有S-腺苷-L-蛋氨酸，腐胺，焦磷酸铁，N-乙酰氨基葡糖胺，腐胺，对氨基苯甲酸，L-半胱氨酸和L-谷氨酰胺的厄尔盐的基本必需培养基。马血清。在31摄氏度的室内空气中孵育。培养基的pH值从8.8开始，并随着细胞的生长上升到大约等于9。根据以中等密度接种的培养物获得的生长曲线计算的加倍时间为35至65小时。使用低密度接种物时，加倍时间减少到19小时。染色涂片和透射电子显微照片中培养的生物的形态是卡氏疟原虫的形态，而P氏癌特异性mAb标记了培养物。培养的生物体对免疫抑制的老鼠具有感染力，可以冷冻保存并用于重新开始培养。

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