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日本血吸虫虫体和细胞的体外培养。

In vitro cultivation of Schistosoma japonicum-parasites and cells.

机构信息

State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, Hubei, PR China.

出版信息

Biotechnol Adv. 2013 Dec;31(8):1722-37. doi: 10.1016/j.biotechadv.2013.09.003. Epub 2013 Sep 24.

Abstract

Schistosomiasis is a serious parasitic zoonosis caused by blood-dwelling flukes of the genus Schistosoma. Understanding functions of genes and proteins of this parasite is important for uncovering this pathogen's complex biology, which will provide valuable information to design new strategies for schistosomiasis control. Effective applications of molecular tools reported to investigate schistosome gene function, such as inhibitor studies and transgenesis, rely on the developments of in vitro cultivation system of this parasite and cells. Besides the in vitro culture studies dealing with Schistosoma mansoni, there are also numerous excellent studies about the in vitro cultivation of Schistosoma japonicum, which were performed by Chinese researchers and published in Chinese journals. Nearly every stage of the life-cycle of S. japonicum, including miracidia, mother sporocysts, cercariae, schistosomula, and egg-laying adult worms, was employed for developing in vitro cultivation methods, being accompanied by the introduction of several media and supplements that helped to improve culture conditions. It was not only possible to generate mother sporocysts from miracidia in vitro, but also to obtain adult worms from cercariae through in vitro cultivation. The main obstacles to complete the life cycle of S. japonicum in the lab are the transition from mother sporocysts to cercariae, and the production of fertilized and completely developed eggs by adult worms generated in vitro. With regard to cells from S. japonicum, besides established isolation protocols and morphological observations, media optimizations were conducted by using different chemical reagents, biological supplements and physical treatment. Among these, mutagens like N-methyl-N-nitro-N-nitrosoguanidine and the addition of extracellular matrix were found to be able to induce mitogenic activities. Although enzyme activities or the level of silver-stained nucleolar region associated protein in cultured cells indicated still suboptimal conditions, the achievements made point to the possibility of reaching the aim of establishing cell lines for S. japonicum. Both the improvements of the in vitro culture of larval and adult worms of S. japonicum as well as the access of cells of this parasite provide excellent advances for research on this important parasite in the future.

摘要

血吸虫病是一种由血吸虫属的血吸吸虫引起的严重寄生虫病。了解该寄生虫的基因和蛋白质功能对于揭示其复杂的生物学特性非常重要,这将为设计新的血吸虫病控制策略提供有价值的信息。已经报道了许多有效的分子工具应用于研究血吸虫基因功能,如抑制剂研究和转基因,这些都依赖于该寄生虫的体外培养系统和细胞的发展。除了涉及曼氏血吸虫的体外培养研究外,还有许多关于日本血吸虫的体外培养的优秀研究,这些研究是由中国研究人员在中文期刊上发表的。日本血吸虫的生命周期的几乎每个阶段,包括毛蚴、母胞蚴、尾蚴、童虫和产卵成虫,都被用于开发体外培养方法,同时引入了几种培养基和补充剂来改善培养条件。不仅可以在体外从毛蚴中产生母胞蚴,而且可以通过体外培养从尾蚴中获得成虫。在实验室中完成日本血吸虫完整生命周期的主要障碍是从母胞蚴到尾蚴的转变,以及体外培养产生的成虫产生受精和完全发育的卵。关于日本血吸虫的细胞,除了已建立的分离方案和形态学观察外,还通过使用不同的化学试剂、生物补充剂和物理处理进行了培养基优化。其中,发现诱变剂如 N-甲基-N-亚硝基-N-亚硝基胍和细胞外基质的添加能够诱导有丝分裂活性。尽管培养细胞的酶活性或银染核仁相关蛋白水平表明仍然存在不理想的条件,但所取得的成就表明有可能达到建立日本血吸虫细胞系的目标。日本血吸虫幼虫和成虫的体外培养的改进以及该寄生虫细胞的获得都为未来对这种重要寄生虫的研究提供了极好的进展。

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