反映囊胚期的滋养外胚层干细胞捕获方案。

Protocol for capturing trophectoderm stem cells reflecting the blastocyst stage.

作者信息

Seong Jinwoo, Rivron Nicolas C

机构信息

Institute of Molecular Biotechnology of the Austrian Academy of Sciences, Vienna Biocenter, 1030 Vienna, Austria.

出版信息

STAR Protoc. 2023 Mar 16;4(2):102151. doi: 10.1016/j.xpro.2023.102151.

DOI:10.1016/j.xpro.2023.102151

PMID:36930647

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10031533/

Abstract

Classically, culturing mouse blastocysts with FGF4/TGF-β1, two epiblast-secreted inducers, allows for deriving trophoblast stem cells that comprise fluctuating subpopulations reflecting both pre- and post-implantation stages. However, a more complete combination of inducers (adding LPA, IL11, BMP7, Activin A, 8-Br-cAMP) captures trophectoderm stem cells with enhanced transcriptomic similarity to the blastocyst trophectoderm and self-renewal, reduced differentiation. Also, the complete combination of inducers increased potential to form blastoids and to instruct decidualization in utero, thus better reflecting the blastocyst. For complete details on the use and execution of this protocol, please refer to Seong et al..

摘要

传统上，用两种外胚层分泌的诱导因子FGF4/TGF-β1培养小鼠囊胚，可以获得滋养层干细胞，这些干细胞包含反映着床前和着床后阶段的波动亚群。然而，更完整的诱导因子组合（添加LPA、IL11、BMP7、激活素A、8-溴-cAMP）捕获的滋养外胚层干细胞与囊胚滋养外胚层具有更高的转录组相似性，自我更新能力增强，分化减少。此外，诱导因子的完整组合增加了形成胚状体和在子宫内诱导蜕膜化的潜力，从而更好地反映囊胚。有关本方案使用和实施的完整详细信息，请参考Seong等人的研究。