Embryo Models In Vitro Methods and Protocols

This collection explores the use of pluripotent cells to generate early embryo-like structures in vitro, with the resultant 3D embryo-like structures in vitro phenocopying many of the developmental landmarks and 3D architectures seen normally in vivo. The protocols gathered herein cover diverse aspects of the topic and exemplify some of the key developments and improvements in the field. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Generation of Human Blastoids from Naïve Pluripotent Stem Cells.- Human Pre-Gastrulation Embryo Culture in 3D Condition.- Protocol for the Generation of Human EPS-Blastoids Using a Three-Dimensional Two-Step Induction System.- Generation of Human Trophoblast Stem Cell-Dependent Placental In Vitro Models.- Workflow for Performing Genetic Manipulation in Human Trophoblast Stem Cells Using CRISPR/Cas9 Technology.- Endometrial Assembloids to Model Human Embryo Implantation In Vitro.- Stem Cell-Derived Microfluidic Amniotic Sac Embryoid (µPASE).- Generation of 3D Trophoblast Organoids from Human Naïve Pluripotent Stem Cells.- Induction of Human Extraembryonic Mesoderm Cells from Naive Pluripotent Stem Cells.- Modeling Human Paraxial Mesoderm Development with Pluripotent Stem Cells.- Generation of Stem Cell-Based Mouse Embryo-Like Structures.- Embryonic Spinal Cord Innervation in Human Trunk Organogenesis Gastruloids: Cardiac Versus Enteric Customization and Beyond.- Use of Epigenetic Cues and Mechanical Stimuli to Generate Blastocyst-Like Structures from Mammalian Skin Dermal Fibroblasts.- Directed Differentiation of Human Pluripotent Stem Cells to Cytotrophoblast and Syncytiotrophoblast.- Single-Cell mRNA-sncRNA Co-Sequencing of Preimplantation Embryos.- Evaluation of Stem-Cell Embryo Models by Integration with a Human Embryo Single-Cell Transcriptome Atlas.- Inferring Gene Regulatory Networks and Predicting the Effect of Gene Perturbations via IQCELL.- ShapeMetrics: A 3D Cell Segmentation Pipeline for Single-Cell Spatial Morphometric Analysis.- Guinea Pig Preimplantation Embryos: Generation, Collection, and Immunofluorescence.- Manual Dissociation of Mammalian Preimplantation Embryos for Single-Cell Genomics.- Whole-Mount RNA, Single Molecule RNA (smRNA), and DNA Fluorescence In Situ Hybridization (FISH) in Mammalian Embryos.