Chick embryo tracheal organ: a new and effective in vitro culture model for Cryptosporidium baileyi.
Chick embryo tracheal organ: a new and effective in vitro culture model for Cryptosporidium baileyi.
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DOI：
10.1016/j.vetpar.2012.03.049
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发表时间：
2012-09
影响因子：
2.6
通讯作者：
Su-mei Zhang;Fuchun Jian;Guanghui Zhao;Lei Huang;Longxian Zhang;Changshen Ning;Rongjun Wang;Meng Qi;Lihua Xiao
中科院分区：
文献类型：
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作者：
Su-mei Zhang;Fuchun Jian;Guanghui Zhao;Lei Huang;Longxian Zhang;Changshen Ning;Rongjun Wang;Meng Qi;Lihua Xiao
用鸡胚气管器(TOCs)培养贝氏隐孢子虫卵囊或子孢子。将I组约5×104个子孢子和卵囊混合物，II组5×105个、III组1×106、IV组2×106个纯化子孢子分别接种于RPMI1640添加5%热灭活胎牛血清的鸡胚气管环中，在40℃、5%CO2的24孔培养板中每孔培养。4个试验组(I-IV组)的气管环被白纹伊蚊成功感染，并在光镜和电子显微镜下观察到不同阶段的寄生虫。寄生虫感染和细胞学改变早在PI 72h时就可观察到。隐孢子虫附着在气管上皮细胞的边缘，其中I组的寄生虫数量多于II、III和IV组，并可见中度的核肿胀和染色质边集，上皮细胞核的正常垂直方向和基底位置几乎消失。经TOCS传代的白纹伊蚊在气管内接种前表现出与寄生虫相似的免疫力和分子特征。这些结果表明，鸡胚气管器官是一种新的、有效的白纹伊蚊和其他呼吸道病原体的体外培养模型。
In the present study, chick embryo tracheal organ (TOCs) was used to cultivate oocysts or sporozoites of Cryptosporidium baileyi. Approximately 5×104sporozoites and oocysts mixture for group I; 5×105, 1×106, 2×106purified sporozoites for group II, group III and group IV, respectively, were inoculated into respective chick embryo tracheal rings maintained in RPMI 1640 supplemented with 5% heat-inactivated FBS, and cultivated in each well of the 24-well culture plate at 40°C and 5% CO2. The tracheal rings in four experimental groups (I–IV) were successfully infected with C. baileyi, and different stages of parasites were also observed under light and electron microscopy. Parasite infection and cytological alterations were noted as early as PI 72h. The Cryptosporidium were seen attached to the edge of the tracheal epithelium, with more number of parasites in group I than that in group II, group III and group IV. The moderate nuclear swelling and chromatin margination were also detected, and the normal vertical orientation and basilar location of the nuclei of the epithelial cells were almost lost. C. baileyi that has been passed by TOCs exhibited similar immunity and molecular features with parasites before intratracheal inoculation. These results suggest that chick embryo tracheal organ is a new and effective in vitro culture model for C. baileyi and other respiratory pathogens.