Scalable cryopreservation of infectious Cryptosporidium hominis oocysts by vitrification.
Scalable cryopreservation of infectious Cryptosporidium hominis oocysts by vitrification.
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DOI：
10.1371/journal.ppat.1011425
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发表时间：
2023-06
期刊：
影响因子：
6.7
通讯作者：
中科院分区：
文献类型：
--
作者：
人隐孢子虫是发展中国家儿童腹泻的一个严重原因。治疗学的发展受到主要技术障碍的阻碍，包括缺乏冷冻保存和简单的培养方法。这影响了用于研究和人类挑战研究的传染性寄生虫卵囊的优化/标准化单一来源的可用性。人类C。人TU 502分离株目前仅在一个实验室中在无菌仔猪中繁殖，这限制了获得卵囊。简化的冷冻保存可以建立一个生物库，作为研究和分发给其他需要C。人类本文报道了C.人型TU 502卵囊，使用专门设计的体积为100 μL的标本容器进行玻璃化冷冻。解冻的卵囊表现出约70%的活力，在无菌仔猪中具有稳健的脱囊和100%的感染率。优化/标准化的卵囊来源的可用性可以通过使更广泛地获得生物标本来简化药物和疫苗评价。我们在这里报告的第一个成功的方法冷冻保存隐孢子虫的人寄生虫，一个最常见的感染性原因的儿科腹泻在发展中国家。这是重要的，因为缺乏这种寄生虫的冷冻保存方法阻碍了科学和临床研究的进展。在缺乏这些方法的情况下，C.必须通过在无菌仔猪中传代来周期性地维持人。因此，这种独特的分离物的冷冻保存是增加研究的共享和可用性并防止灾难性损失的优先事项。冷冻保存也是必要的，以促进人类挑战研究和临床试验，以评估治疗。在这里，我们成功地冷冻保存的寄生虫玻璃化快速冷却速度达到专门设计的标本容器。我们发现，解冻的寄生虫保留生活力和感染性的gnotobiotic仔猪。值得注意的是，这些方法使我们能够建立一个C。参考分离株TU 502人卵囊。生物库优化/标准化卵囊制剂的可用性可能有助于药物和疫苗评价。
Cryptosporidium hominis is a serious cause of childhood diarrhea in developing countries. The development of therapeutics is impeded by major technical roadblocks including lack of cryopreservation and simple culturing methods. This impacts the availability of optimized/standardized singular sources of infectious parasite oocysts for research and human challenge studies. The human C. hominis TU502 isolate is currently propagated in gnotobiotic piglets in only one laboratory, which limits access to oocysts. Streamlined cryopreservation could enable creation of a biobank to serve as an oocyst source for research and distribution to other investigators requiring C. hominis. Here, we report cryopreservation of C. hominis TU502 oocysts by vitrification using specially designed specimen containers scaled to 100 μL volume. Thawed oocysts exhibit ~70% viability with robust excystation and 100% infection rate in gnotobiotic piglets. The availability of optimized/standardized sources of oocysts may streamline drug and vaccine evaluation by enabling wider access to biological specimens. We report here on the first successful method of cryopreservation of Cryptosporidium hominis parasites, one of the most common infectious causes of pediatric diarrhea in the developing world. This is significant as lack of cryopreservation methods for this species of parasite hinders progress of scientific and clinical research. In the absence of these methods, a reference isolate of C. hominis must be maintained periodically by passage in gnotobiotic piglets. Cryopreservation of this unique isolate is therefore a priority to increase sharing and availability for research, and to prevent catastrophic loss. Cryopreservation is also necessary to facilitate human challenge studies and clinical trials for the evaluation of therapeutics. Here, we successfully cryopreserved the parasite by vitrification using rapid cooling rates achieved in specially designed specimen containers. We showed that the thawed parasite retained viability and infectivity in gnotobiotic piglets. Notably these methods allowed us to establish a biobank of C. hominis oocysts of reference isolate TU502. The availability of biobanked optimized/standardized oocyst preparations may facilitate drug and vaccines evaluation.
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影响因子：
1.3
作者：
Basso, W;Venturini, L;Dubey, JP
通讯作者：
Dubey, JP
影响因子：
3.3
作者：
Ifeonu, Olukemi O.;Chibucos, Marcus C.;Silva, Joana C.
通讯作者：
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DOI：
10.1016/s2214-109x(18)30283-3
发表时间：
2018-07
期刊：
The Lancet. Global health
影响因子：
--
作者：
Khalil IA;Troeger C;Rao PC;Blacker BF;Brown A;Brewer TG;Colombara DV;De Hostos EL;Engmann C;Guerrant RL;Haque R;Houpt ER;Kang G;Korpe PS;Kotloff KL;Lima AAM;Petri WA Jr;Platts-Mills JA;Shoultz DA;Forouzanfar MH;Hay SI;Reiner RC Jr;Mokdad AH
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影响因子：
16.6
作者：
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1.3
作者：
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通讯作者：
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