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Embryonic Stem Cell Protocols

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Cover of 'Embryonic Stem Cell Protocols'

Table of Contents

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    Book Overview
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    Chapter 126 Osteogenic Differentiation from Embryonic Stem Cells.
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    Chapter 152 Derivation of Neural Precursor Cells from Human Embryonic Stem Cells for DNA Methylomic Analysis.
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    Chapter 207 Acquiring Ground State Pluripotency: Switching Mouse Embryonic Stem Cells from Serum/LIF Medium to 2i/LIF Medium.
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    Chapter 208 From Naive to Primed Pluripotency: In Vitro Conversion of Mouse Embryonic Stem Cells in Epiblast Stem Cells.
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    Chapter 209 Generation of Embryonic Stem Cells in Rabbits.
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    Chapter 210 Applying Shear Stress to Pluripotent Stem Cells
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    Chapter 211 A Simple and Efficient Method of Slow Freezing for Human Embryonic Stem Cells and Induced Pluripotent Stem Cells.
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    Chapter 212 A Concise Protocol for siRNA-Mediated Gene Suppression in Human Embryonic Stem Cells
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    Chapter 213 Generation of a Knockout Mouse Embryonic Stem Cell Line Using a Paired CRISPR/Cas9 Genome Engineering Tool.
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    Chapter 214 Expanding the Utility of FUCCI Reporters Using FACS-Based Omics Analysis
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    Chapter 215 Generating Inner Ear Organoids from Mouse Embryonic Stem Cells.
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    Chapter 216 26S and PA28-20S Proteasome Activity in Cytosolic Extracts from Embryonic Stem Cells
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    Chapter 217 Pancreatic Differentiation from Murine Embryonic Stem Cells
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    Chapter 218 In Vitro Differentiation of Embryonic Stem Cells into Hematopoietic Lineage: Towards Erythroid Progenitor's Production.
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    Chapter 219 Differentiation of Adipocytes in Monolayer from Mouse Embryonic Stem Cells
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    Chapter 220 Generation and Purification of Definitive Endoderm Cells Generated from Pluripotent Stem Cells
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    Chapter 221 Gene Transfer into Pluripotent Stem Cells via Lentiviral Transduction
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    Chapter 226 Delivering Antisense Morpholino Oligonucleotides to Target Telomerase Splice Variants in Human Embryonic Stem Cells.
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    Chapter 228 Maintenance, Transgene Delivery, and Pluripotency Measurement of Mouse Embryonic Stem Cells.
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    Chapter 229 Limbal Stromal Tissue Specific Stem Cells and Their Differentiation Potential to Corneal Epithelial Cells.
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    Chapter 230 In Vitro Differentiation of Pluripotent Stem Cells into Functional β Islets Under 2D and 3D Culture Conditions and In Vivo Preclinical Validation of 3D Islets.
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    Chapter 231 Generation of Corneal Keratocytes from Human Embryonic Stem Cells.
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    Chapter 232 Embryonic Stem Cell Protocols
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    Chapter 233 Analysis of mRNA Translation Rate in Mouse Embryonic Stem Cells.
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    Chapter 234 Methods for Derivation of Multipotent Neural Crest Cells Derived from Human Pluripotent Stem Cells.
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    Chapter 235 Dopaminergic Differentiation of Human Embryonic Stem Cells on PA6-Derived Adipocytes
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    Chapter 253 Maximizing Clonal Embryonic Stem Cell Derivation by ERK Pathway Inhibition.
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    Chapter 254 Resolving Heterogeneity: Fluorescence-Activated Cell Sorting of Dynamic Cell Populations from Feeder-Free Mouse Embryonic Stem Cell Culture.
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    Chapter 255 Imaging Pluripotency: Time-Lapse Analysis of Mouse Embryonic Stem Cells
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    Chapter 270 Selection of Antibodies Interfering with Cell Surface Receptor Signaling Using Embryonic Stem Cell Differentiation.
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    Chapter 271 Embryonic Stem Cell Protocols
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    Chapter 272 Definitive Endoderm Differentiation of Human Embryonic Stem Cells Combined with Selective Elimination of Undifferentiated Cells by Methionine Deprivation
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    Chapter 313 Erratum to: Resolving Heterogeneity: Fluorescence-Activated Cell Sorting of Dynamic Cell Populations from Feeder-Free Mouse Embryonic Stem Cell Culture
Attention for Chapter 270: Selection of Antibodies Interfering with Cell Surface Receptor Signaling Using Embryonic Stem Cell Differentiation.
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Chapter title
Selection of Antibodies Interfering with Cell Surface Receptor Signaling Using Embryonic Stem Cell Differentiation.
Chapter number 270
Book title
Embryonic Stem Cell Protocols
Published in
Methods in molecular biology, June 2015
DOI 10.1007/7651_2015_270
Pubmed ID
Book ISBNs
978-1-4939-2953-5, 978-1-4939-2954-2
Authors

Melidoni, Anna N, Dyson, Michael R, McCafferty, John, Melidoni, Anna N., Dyson, Michael R., Anna N. Melidoni, Michael R. Dyson, John McCafferty

Abstract

Antibodies able to bind and modify the function of cell surface signaling components in vivo are increasingly being used as therapeutic drugs. The identification of such "functional" antibodies from within large antibody pools is, therefore, the subject of intense research. Here we describe a novel cell-based expression and reporting system for the identification of functional antibodies from antigen-binding populations preselected with phage display. The system involves inducible expression of the antibody gene population from the Rosa-26 locus of embryonic stem (ES) cells, followed by secretion of the antibodies during ES cell differentiation. Target antigens are cell-surface signaling components (receptors or ligands) with a known effect on the direction of cell differentiation (FGFR1 mediating ES cell exit from self renewal in this particular protocol). Therefore, inhibition or activation of these components by functional antibodies in a few elite clones causes a shift in the differentiation outcomes of these clones, leading to their phenotypic selection. Functional antibody genes are then recovered from positive clones and used to produce the purified antibodies, which can be tested for their ability to affect cell fates exogenously. Identified functional antibody genes can be further introduced in different stem cell types. Inducible expression of functional antibodies has a temporally controlled protein-knockdown capability, which can be used to study the unknown role of the signaling pathway in different developmental contexts. Moreover, it provides a means for control of stem cell differentiation with potential in vivo applications.

Mendeley readers

The data shown below were compiled from readership statistics for 14 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
United States 1 7%
Germany 1 7%
Unknown 12 86%

Demographic breakdown

Readers by professional status Count As %
Researcher 5 36%
Student > Master 3 21%
Student > Bachelor 3 21%
Librarian 1 7%
Unknown 2 14%
Readers by discipline Count As %
Agricultural and Biological Sciences 5 36%
Biochemistry, Genetics and Molecular Biology 4 29%
Pharmacology, Toxicology and Pharmaceutical Science 1 7%
Computer Science 1 7%
Medicine and Dentistry 1 7%
Other 0 0%
Unknown 2 14%