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Large-Quantity Production of Chicken Embryo Tracheal Organ Cultures and Use in Virus and Mycoplasma Studies

机译:鸡胚气管器官培养物的大量生产及其在病毒和支原体研究中的应用

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摘要

Chicken tracheal organ cultures were made from embryos which were 19 to 20 days old. Transversely cut rings of trachea were placed in screw-capped tissue-culture tubes with Eagle's-N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid (HEPES) medium and incubated in roller drums. The method had advantages over other organ culture systems in that these cultures were prepared in numbers similar to conventional tissue cultures, ciliary activity was quickly and accurately evaluated, and contamination occurred less frequently than with organ cultures in petri dishes. Ciliary activity persisted for at least 1 month when the medium was changed at 5-to 7-day intervals and for 10 to 15 days without a change. Infectious bronchitis virus stopped ciliary movement, and this effect was used as a basis for titrating the virus and for determining the neutralizing capacity of immune mouse ascitic fluid. Twenty-four Mycoplasma strains were tested. Organisms of 17 strains, both avian and mammalian, multiplied in the organ cultures, and 7 strains, belonging to the species M. gallisepticum and M. mycoides var. capri, inhibited ciliary activity.
机译:鸡气管器官培养物是由19至20天大的胚胎制成的。将横向切开的气管环放入装有Eagle's-N-2-羟乙基哌嗪-N'-2-乙烷磺酸(HEPES)培养基的螺旋盖组织培养管中,并在转鼓中孵育。与其他器官培养系统相比,该方法具有优势,因为这些培养物的制备数量与常规组织培养物相似,可以快速准确地评估睫毛活动,并且与培养皿中的器官培养相比,污染发生的频率更低。当以5到7天的间隔更换培养基时,睫毛活动持续至少1个月,而没有变化则持续10到15天。传染性支气管炎病毒使纤毛运动停止,该作用被用作滴定病毒和确定免疫小鼠腹水的中和能力的基础。测试了二十四支支原体菌株。禽类和哺乳动物中有17个菌株的生物在器官培养物中倍增,而属于鸡毒支原体和霉菌支原体的7种菌株。卡普里岛,抑制睫毛活动。

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