Chimeric 3D-gastruloids – a versatile tool for studies of mammalian peri-gastrulation development
Chimeric 3D-gastruloids – a versatile tool for studies of mammalian peri-gastrulation development
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嵌合 3D-原肠胚样蛋白——研究哺乳动物原肠胚形成期间发育的多功能工具
DOI：
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
S. Arnold
中科院分区：
文献类型：
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作者：
Alexandra E. Wehmeyer;Katrin M. Schüle;Alexandra Conrad;Chiara M. Schröder;S. Probst;S. Arnold
干细胞衍生的3d类原肠细胞显示出显著的自组织能力，并概括了哺乳动物原肠形成阶段发育的许多方面。胃原体可以快速生成，并提供几个实验优势，如可扩展性、可观察性和可操作性。在这里，我们提出了利用小鼠胚胎干细胞（mESCs）的功能遗传学来产生嵌合类胃原体的方法，以进一步扩大小鼠3d类胃原体的实验效力。在嵌合的类胃原体中，荧光标记的含有诱导基因表达或功能缺失等位基因的不同基因型细胞与野生型细胞结合。我们在携带Tbx转录因子Brachyury纯合缺失或Eomes诱导表达的mESCs嵌合原肠样细胞中展示了这种实验方法。由此产生的嵌合性胃原体分别概括了报道的Eomes和Brachyury功能，如指示心脏命运和促进后轴向伸展。此外，嵌合类原肠细胞揭示了以前未被认识到的表型，如brachyury缺失细胞对内胚层的组织分选偏好，以及brachyury缺失对胚胎轴上wnt3a模式的非自主性影响，证明了嵌合类原肠细胞作为哺乳动物原肠形成研究的有效工具的一些优势。
Stem cell-derived 3D-gastruloids show a remarkable capacity of self-organisation and recapitulate many aspects of gastrulation stage mammalian development. Gastruloids can be rapidly generated and offer several experimental advantages, such as scalability, observability, and accessibility for manipulation. Here, we present approaches to further expand the experimental potency of murine 3D-gastruloids by utilizing functional genetics in mouse embryonic stem cells (mESCs) to generate chimeric gastruloids. In chimeric gastruloids fluorescently labelled cells of different genotypes harbouring inducible gene-expression, or loss-of-function alleles, are combined with wildtype cells. We showcase this experimental approach in chimeric gastruloids of mESCs carrying homozygous deletions of the Tbx transcription factors Brachyury, or inducible expression of Eomes. Resulting chimeric gastruloids recapitulate reported Eomes and Brachyury functions, such as instructing cardiac fate and promoting posterior axial extension, respectively. Additionally, chimeric gastruloids revealed previously unrecognized phenotypes such as tissue sorting preference of Brachyury-deficient cells to endoderm, and cell non-autonomous effects of Brachyury-deficiency on Wnt3a-patterning along the embryonic axis, demonstrating some of the advantages of chimeric gastruloids as efficient tool for studies of mammalian gastrulation.
影响因子：
23.9
作者：
Rossi G;Broguiere N;Miyamoto M;Boni A;Guiet R;Girgin M;Kelly RG;Kwon C;Lutolf MP
通讯作者：
Lutolf MP
影响因子：
11.8
作者：
Martin, Benjamin L.;Kimelman, David
通讯作者：
Kimelman, David