培养基和培养方式会改变猪结肠单层的细胞组成和屏障完整性。

IF 3.6 Q2 MEDICINE, RESEARCH & EXPERIMENTAL Tissue Barriers Pub Date : 2024-04-02 Epub Date: 2023-06-21 DOI:10.1080/21688370.2023.2222632
Alicia M Barnett, Jane A Mullaney, Warren C McNabb, Nicole C Roy
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引用次数: 0

摘要

肠道类器官技术彻底改变了我们的体外细胞培养方法,部分原因是其三维结构在细胞组成和结构方面更像原生组织。因此,类器官正成为进行肠上皮细胞研究的新黄金标准。遗憾的是,有机体本来具有三维几何形状的优势,但却无法轻松进入顶端上皮细胞,这在研究饮食或微生物成分与宿主组织之间的相互作用时是一个主要限制。为了克服这一问题,我们开发了猪结肠源单层细胞,在可渗透的 Transwell 插入物和经过组织培养处理的聚苯乙烯板上进行培养。我们发现,播种密度和培养形式改变了编码特定细胞类型(干细胞、结肠细胞、鹅口疮细胞和肠内分泌细胞)和屏障成熟(紧密连接)标志物的基因的表达。此外,我们还发现,培养基配方的改变会改变结肠组织及其衍生单层细胞的细胞组成,从而使培养物的分化表型越来越接近其原生组织的表型。
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Culture media and format alter cellular composition and barrier integrity of porcine colonoid-derived monolayers.

Intestinal organoid technology has revolutionized our approach to in vitro cell culture due in part to their three-dimensional structures being more like the native tissue from which they were derived with respect to cellular composition and architecture. For this reason, organoids are becoming the new gold standard for undertaking intestinal epithelial cell research. Unfortunately, their otherwise advantageous three-dimensional geometry prevents easy access to the apical epithelium, which is a major limitation when studying interactions between dietary or microbial components and host tissues. To overcome this problem, we developed porcine colonoid-derived monolayers cultured on both permeable Transwell inserts and tissue culture treated polystyrene plates. We found that seeding density and culture format altered the expression of genes encoding markers of specific cell types (stem cells, colonocytes, goblets, and enteroendocrine cells), and barrier maturation (tight junctions). Additionally, we found that changes to the formulation of the culture medium altered the cellular composition of colonoids and of monolayers derived from them, resulting in cultures with an increasingly differentiated phenotype that was similar to that of their tissue of origin.

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来源期刊
Tissue Barriers
Tissue Barriers MEDICINE, RESEARCH & EXPERIMENTAL-
CiteScore
6.60
自引率
6.50%
发文量
25
期刊介绍: Tissue Barriers is the first international interdisciplinary journal that focuses on the architecture, biological roles and regulation of tissue barriers and intercellular junctions. We publish high quality peer-reviewed articles that cover a wide range of topics including structure and functions of the diverse and complex tissue barriers that occur across tissue and cell types, including the molecular composition and dynamics of polarized cell junctions and cell-cell interactions during normal homeostasis, injury and disease state. Tissue barrier formation in regenerative medicine and restoration of tissue and organ function is also of interest. Tissue Barriers publishes several categories of articles including: Original Research Papers, Short Communications, Technical Papers, Reviews, Perspectives and Commentaries, Hypothesis and Meeting Reports. Reviews and Perspectives/Commentaries will typically be invited. We also anticipate to publish special issues that are devoted to rapidly developing or controversial areas of research. Suggestions for topics are welcome. Tissue Barriers objectives: Promote interdisciplinary awareness and collaboration between researchers working with epithelial, epidermal and endothelial barriers and to build a broad and cohesive worldwide community of scientists interesting in this exciting field. Comprehend the enormous complexity of tissue barriers and map cross-talks and interactions between their different cellular and non-cellular components. Highlight the roles of tissue barrier dysfunctions in human diseases. Promote understanding and strategies for restoration of tissue barrier formation and function in regenerative medicine. Accelerate a search for pharmacological enhancers of tissue barriers as potential therapeutic agents. Understand and optimize drug delivery across epithelial and endothelial barriers.
期刊最新文献
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