Department of Molecular and Cellular Biology, University of Guelph, 50 Stone Road East, Guelph, Ontario N1G 2W1, Canada.
J Microbiol Methods. 2013 Nov;95(2):167-74. doi: 10.1016/j.mimet.2013.08.008. Epub 2013 Aug 27.
In vitro gut models provide several advantages over in vivo models for the study of the human gut microbiota. However, because communities developed in these models are inevitably simplified simulations of the in vivo environment, it is necessary to broadly define the differences between in vitro consortia and the communities from which they are derived. In this study we characterized microbial community development in a twin-vessel single-stage chemostat model of the human distal gut ecosystem using both gel (Denaturing Gradient Gel Electrophoresis) and phylogenetic microarray (Human Intestinal Tract Chip) based techniques. Five different sets of twin-vessels were inoculated with feces from three different healthy adult donors and allowed to reach steady state compositions. We found that twin-vessel single-stage chemostats could develop and maintain stable, diverse, and reproducible communities that reach steady state compositions in all five runs by at most 36 days post-inoculation. As noted in other in vitro studies, steady state communities were enriched in Bacteroidetes but not Clostridium cluster XIVa, Bacilli or other Firmicutes relative to the fecal inocula. Communities developed within this model had higher within-run reproducibility than between-run repeatability when using consecutive fecal donations. Both fecal inocula and steady state chemostat communities seeded with feces from different donors had distinct compositions. We conclude that twin-vessel single-stage chemostat models represent a valid simulation of the human distal gut environment and can support complex, representative microbial communities ideal for experimental manipulation.
在研究人类肠道微生物群时,体外肠道模型相对于体内模型具有许多优势。然而,由于这些模型中开发的群落不可避免地是对体内环境的简化模拟,因此有必要广泛定义体外群落与它们衍生的群落之间的差异。在这项研究中,我们使用凝胶(变性梯度凝胶电泳)和基于系统发育的微阵列(人类肠道芯片)技术来描述人类远端肠道生态系统双容器单级恒化器模型中微生物群落的发展。用来自三个不同健康成人供体的粪便接种五组不同的双容器,并允许其达到稳定状态的组成。我们发现,双容器单级恒化器可以在最多 36 天的接种后达到稳定状态的组成,从而发展和维持稳定,多样且可重复的群落。与粪便接种物相比,正如其他体外研究中指出的那样,稳态群落富含拟杆菌门,但不富含梭菌簇 XIVa、芽孢杆菌或其他厚壁菌门。在该模型中,当使用连续粪便供体时,群落的内部运行重复性高于运行间重复性。来自不同供体的粪便接种的粪便和稳定的恒化器群落具有不同的组成。我们得出结论,双容器单级恒化器模型代表了对人类远端肠道环境的有效模拟,并且可以支持复杂,具有代表性的微生物群落,非常适合实验操作。