Continuous axenic cultivation of Pneumocystis carinii.
Continuous axenic cultivation of Pneumocystis carinii.
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卡氏肺囊虫的连续无菌培养。
DOI：
10.1073/pnas.96.5.2402
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发表时间：
1999
影响因子：
11.1
通讯作者：
ClarksonJr,AB
中科院分区：
文献类型：
--
作者：
Merali,S;Frevert,U;Williams,JH;Chin,K;Bryan,R;ClarksonJr,AB
实现了卡氏肺孢子虫的连续无菌培养。使用的培养容器允许频繁更换培养基，而不会干扰附着在胶原蛋白涂层多孔膜上生长的微生物。生长培养基基于含有Earle盐的最小必需培养基，补充有S-腺苷-L-甲硫氨酸、腐胺、焦磷酸铁、N-乙酰葡糖胺、腐胺、对氨基苯甲酸、L-半胱氨酸和L-谷氨酰胺以及马血清。孵育在31°C的室内空气中进行。培养基的pH值从8.8开始，随着细胞的生长上升到8.9。从以中等密度接种的培养物获得的生长曲线计算的倍增时间范围为35至65小时。使用低密度接种物，倍增时间缩短至19小时。在染色涂片和透射电镜照片中培养的生物体的形态是P. carinii，和P.卡氏体特异性mAb标记培养材料。培养的微生物对免疫抑制大鼠具有感染性，可冷冻储存并用于重新启动培养。
Continuous axenic culture ofPneumocystis cariniihas 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 membrane. The growth medium is based on Minimal Essential Medium with Earle’s salt supplemented withS-adenosyl-l-methionine, putrescine, ferric pyrophosphate,N-acetyl glucosamine, putrescine,p-aminobenzoic acid,l-cysteine andl-glutamine, and horse serum. Incubation is in room air at 31°C. The pH of the medium begins at 8.8 and rises to ≈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 19 hours. The morphology of cultured organisms in stained smears and in transmission electron micrographs is that ofP. carinii, andP. carinii-specific mAbs label the cultured material. Cultured organisms are infective for immunosuppressed rats and can be stored frozen and used to reinitiate culture.