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New advances in the in-vitro culture of Dientamoeba fragilis

机译：脆弱Dientamoeba体外培养的新进展

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Dientamoeba fragilis is an intestinal protozoan in humans that is commonly associated with diarrhoea and other gastrointestinal complaints. Studies conducted to investigate the biology of this parasite are limited by methods for in vitro cultivation. The objective of this study was to improve a biphasic culture medium, based on the Loeffler's slope, by further supplementation in order to increase the yield of trophozoites in culture. The current in vitro culture of D. fragilis is a xenic culture with a mix of bacteria. Three different liquid overlays were evaluated including Earle's balanced salt solution (EBSS), PBS and Dulbecco's modified PBS (DPBS), for their ability to support the in vitro growth of D. fragilis trophozoites. Out of these 3 overlays EBSS gave the highest increase in the trophozoite numbers. The effect of supplementation was analysed by supplementing EBSS with ascorbic acid, ferric ammonium citrate, L-cysteine, cholesterol and alpha-lipoic acid and quantification of in vitro growth by cell counts. A new liquid overlay is here described based upon EBSS supplemented with cholesterol and ferric ammonium citrate that, in conjunction with the Loeffler's slope, supports the growth of D. fragilis trophozoites in vitro. This modified overlay supported a 2-fold increase in the numbers of trophozoite in culture from all 4 D. fragilis isolates tested, when compared to a PBS overlay. These advances enable the harvest of a larger number of trophozoites needed for further studies on this parasite.
机译：脆弱龙眼是人类的肠道原生动物，通常与腹泻和其他胃肠道不适有关。为研究该寄生虫的生物学而进行的研究受到体外培养方法的限制。这项研究的目的是通过进一步补充以提高Lopholer坡度为基础的双相培养基，以增加培养物中滋养体的产量。当前的脆弱拟杆菌的体外培养是混合细菌的异种培养。评估了三种不同的液体覆盖层，包括伯氏平衡盐溶液（EBSS），PBS和Dulbecco修饰的PBS（DPBS），以支持脆弱的D. fragilis滋养体的体外生长。在这三个覆盖层中，EBSS的滋养体数量增加最多。通过向EBSS补充抗坏血酸，柠檬酸铁铵，L-半胱氨酸，胆固醇和α-硫辛酸来分析补充效果，并通过细胞计数定量体外生长。本文描述了一种新的液体覆盖物，其基于EBSS补充了胆固醇和柠檬酸铁铵，再加上Loeffler斜率，可支持脆弱的D. fragilis滋养体在体外的生长。与PBS覆盖层相比，这种改良的覆盖层支持来自所有测试的4种脆弱拟杆菌（D. fragilis）分离物中培养物中滋养体数目的2倍增加。这些进展使得能够收获更多的滋养体，这是对该寄生虫进行进一步研究所需的。

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