小鼠胚胎干细胞扩展培养原肠样细胞的优化方案。

Q4 Biochemistry, Genetics and Molecular Biology

Methods in molecular biology Pub Date : 2025-09-20 DOI:10.1007/7651_2025_666

Marloes Blotenburg, Shruthi Buddharaju, Beril Esin, Peter Zeller

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引用次数: 0

摘要

类胃原体是来源于小鼠胚胎干细胞的三维聚集体，它概括了早期胚胎发生的关键事件。生成类胃原体的方案对聚集条件高度敏感，常常导致变异。在这里，我们提出了一种优化的生成原肠样物质的方法，并通过将其包埋在10%的Matrigel中96小时将其培养延长至聚集后168小时。该方案可以用所有三个胚层的衍生物再生生成原肠样物质，为体外研究原肠样物质的发育过程提供了一个扩展的实验窗口。

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An Optimized Protocol for the Extended Culture of Gastruloids from Mouse Embryonic Stem Cells.

Gastruloids are three-dimensional aggregates, derived from mouse embryonic stem cells, that recapitulate key events of early embryogenesis. Protocols to generate gastruloids are highly sensitive to aggregation conditions, often resulting in variability. Here, we present an optimized method for generating gastruloids and extending their culture up to 168 h post-aggregation by embedding them in 10% Matrigel at 96 h. This protocol enables reproducible generation of gastruloids with derivatives of all three germ layers, providing an extended experimental window to study post-gastrulation developmental processes in vitro.

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来源期刊

Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

2.00

自引率

0.00%

发文量

3536

期刊介绍： For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.