{"title":"From Single Stem Cells to an In Vitro Model of the Post-implantation Human Embryo: A Step-by-Step Guide.","authors":"Seher Ipek Gassaloglu, Monique Pedroza, Berna Sozen","doi":"10.1007/7651_2025_647","DOIUrl":null,"url":null,"abstract":"<p><p>Recent advances in three-dimensional (3D) modeling of post-implantation human embryos using human pluripotent stem cells (hPSCs) have revolutionized our ability to investigate this crucial yet enigmatic stage of development. Here we detail the generation of the human extra-embryoid (hEE), a 3D stem cell-based embryo model that uniquely captures key spatiotemporal events of peri-gastrulation development through the formation and co-development of post-implantation embryonic and extra-embryonic lineages, with high efficiency and robustness across genetic backgrounds. This chapter provides a detailed protocol for generating hEEs in vitro, including guidance on hPSC maintenance, expected cell morphology, troubleshooting strategies, and key culture techniques.</p>","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2025-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Methods in molecular biology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1007/7651_2025_647","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 0
Abstract
Recent advances in three-dimensional (3D) modeling of post-implantation human embryos using human pluripotent stem cells (hPSCs) have revolutionized our ability to investigate this crucial yet enigmatic stage of development. Here we detail the generation of the human extra-embryoid (hEE), a 3D stem cell-based embryo model that uniquely captures key spatiotemporal events of peri-gastrulation development through the formation and co-development of post-implantation embryonic and extra-embryonic lineages, with high efficiency and robustness across genetic backgrounds. This chapter provides a detailed protocol for generating hEEs in vitro, including guidance on hPSC maintenance, expected cell morphology, troubleshooting strategies, and key culture techniques.
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.
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