Embryonic Stem Cell Protocols

by Kursad Turksen (Editor)

Overview

This fully updated volume explores improved and extended prools for embryonic stem cell (ESC) research, provided with the most direct and informative methodologies currently available. The book examines how these models for cell lineage and differentiation studies have continued to mature into a critical research workhorse. Written for the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory prools, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and up-to-date, Embryonic Stem Cell Prools, Fourth Edition serves as an ideal resource for researching mining the depths of the ESC field.

Product Details

ISBN-13:	9781071624364
Publisher:	Springer US
Publication date:	07/26/2022
Series:	Methods in Molecular Biology , #2520
Edition description:	4th ed. 2022
Pages:	363
Product dimensions:	7.01(w) x 10.00(h) x (d)

Table of Contents

Dissecting Molecular Phenotypes through FACS-Based Pooled CRISPR Screens.- Feeder Free Human Embryonic Stem Cell Culture under Defined Culture Conditions.- Rat Induced Pluripotent Stem Cells-Derived Cardiac Myocytes in a Cell Culture Dish.- Transient Induction and Characterization of Mouse Epiblast-Like Cells from Mouse Embryonic Stem Cells.- High-Content Image Analysis of Spatiotemporal Proliferation and Differentiation Patterns in 3D Embryoid Body Differentiation Model.- Derivation of Multipotent Neural Progenitors from Human Embryonic Stem Cells for Cell Therapy and Biomedical Applications.- Feeder Dependent/Independent Mouse Embryonic Stem Cell Culture Prool.- ChIP-qPCR for Polycomb Group Proteins during Neuronal Differentiation of Human Pluripotent Stem Cells.- Directed Differentiation of Human Pluripotent Stem Cells into Inner Ear Organoids.- Glycolytic Profiling of Mouse Embryonic Stem Cells (mESCs).- Replating Prool for Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes.- Hematopoietic Cell Isolation by Antibody-Free Flow Cytometry in the Zebrafish Embryo.- Human Trophectoderm Spheroid Derived from Human Embryonic Stem Cells.- Embryoid Bodies-Based Multilineage Differentiation of Human Embryonic Stem Cells Grown on Feeder-Free Conditions.- A Simple, Rapid, and Cost-effective Method for Loss-of-Function Assays in Pluripotent Cells.- Embryoid Body Formation from Mouse and Human Pluripotent Stem Cells for Transplantation to Study Brain Microenvironment and Cellular Differentiation.- A Modified SMART-Seq Method for Single Cell Transcriptomic Analysis of Embryoid Body Differentiation.- Osteogenic Differentiation from Mouse Embryonic Stem Cells.- Murine Embryonic Stem Cell Culture, Self-Renewal, and Differentiation.- Mouse Embryonic Stem Cell Culture in Serum-Containing or 2i Conditions.- Directed Differentiation of Mouse Embryonic Stem Cells to Mesoderm, Endoderm, and Neuroectoderm Lineages.- Accessing the Human Pluripotent Stem Cell Translatome byPolysome Profiling.- Genome Engineering Human ESCs or iPSCs with Cytosine and Adenine Base Editors.- Proteomic Analysis of Human Neural Stem Cells Differentiation by SWATH-MS.

Editorial Reviews

Reviewer:Bruce A. Fenderson, PhD(Thomas Jefferson University)
Description:Stem cells are characterized by their ability to undergo self-renewal, while retaining the potential for terminal differentiation. There is hope that embryonic stem (ES) cells may someday provide tailor-made replacement cells and tissues for patients with chronic diseases (regenerative medicine). This is part of a two-volume set that describes laboratory protocols and experimental models that are currently used in stem cell research. These books are part of the popular Methods in Molecular Biology series. Volume 1 describes methods for stem cell isolation and characterization. Volume 2 provides examples of differentiation models. The two volumes include 62 chapters on topics ranging from isolation of Medaka ES cells and use of ES cells in embryotoxicity assays, to oligodendrocytes from lineage-selectable ES cells. The primary focus is on cell biology, biochemistry, and molecular biology.
Purpose:According to the editor, the aims of these books are to "complement the original mouse ES cell protocol book...focus on ES cells from non-mouse species...and provide new protocols for the use of ES cells in studies of diverse cell lineages." The editor hopes that these research-oriented protocols will help accelerate the pace of discovery in this field.
Audience:These stem cell protocol books are written for basic science and clinical researchers interested in cellular and developmental biology, biochemistry, and molecular biology. Biomedical researchers interested in bioengineering, gene therapy, oncology, and regenerative medicine will also appreciate the methods described here.
Features:Volume 1 is divided into seven major sections, including sections on signaling in ES cell differentiation, genetic manipulation of ES cells, and tumor-like properties of ES cells. Volume 2 includes chapters purporting to illustrate the conversion of nondescript ES cells to a wide variety of differentiated cell types, including lymphocytes, neurons, endothelial cells, and cardiomyocytes to name a few. Each chapter provides a summary, introduction, detailed step-by-step protocols, helpful notes, and references. The notes section allows the authors to provide rationales, further details, and tips for success. The figures and illustrations are reproduced in black and white. The authors are experts in this field.
Assessment:The major attraction of this two-volume set is the wide range of topics it covers. It is interesting to look at examples of how ES cells are derived from fish, chickens, and other species. An important issue raised in Volume 1 concerns the propensity of ES cells to form tumors in vivo. Clearly, there are major hurdles that must be cleared in order to make cell engineering safe and clinically effective. The protocols are typically organized around specific research questions, and numerous examples of research findings are presented. Thus, in addition to gaining access to methods, the reader benefits from a review of research questions under active investigation. One caveat is that protocols for lineage-directed differentiation need to be considered work in progress. This field is hampered by a lack of definitive markers for differentiated cells. Morphology is insufficient evidence for terminal differentiation and normal physiological function. These books will be a welcome resource for research scientists interested in stem cell biology.