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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 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 title
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.
Chapter number 230
Book title
Embryonic Stem Cell Protocols
Published in
Methods in molecular biology, March 2015
DOI 10.1007/7651_2015_230
Pubmed ID
Book ISBNs
978-1-4939-2953-5, 978-1-4939-2954-2

Bipasha Bose, Sudheer Shenoy P, P Shenoy Sudheer, Bose, Bipasha, Sudheer, P Shenoy


Since the advent of pluripotent stem cells, (embryonic and induced pluripotent stem cells), applications of such pluripotent stem cells are of prime importance. Indeed, scientists are involved in studying the basic biology of pluripotent stem cells, but equal impetus is there to direct the pluripotent stem cells into multiple lineages for cell therapy applications. Scientists across the globe have been successful, to a certain extent, in obtaining cells of definitive endoderm and also pancreatic β islets by differentiating human pluripotent stem cells. Pluripotent stem cell differentiation protocols aim at mimicking in vivo embryonic development. As in vivo embryonic development is a complex process and involves interplay of multiple cytokines, the differentiation protocols also involve a stepwise use of multiple cytokines. Indeed the novel markers for pancreas organogenesis serve as the roadmaps to develop new protocols for pancreatic differentiation from pluripotent stem cells. Earliest developed protocols for pancreas differentiation involved "Nestin selection pathway," a pathway common for both neuronal and pancreatic differentiation lead to the generation of cells that were a combination of cells from neuronal lineage. Eventually with the discovery of hierarchy of β cell transcription factors like Pdx1, Pax4, and Nkx2.2, forced expression of such transcription factors proved successful in converting a pluripotent stem cell into a β cell. Protocols developed almost half a decade ago to the recent ones rather involve stepwise differentiations involving various cytokines and could generate as high as 25 % functional insulin-positive cells in vitro. Most advanced protocols for β islet differentiations from human pluripotent stem cells focused on 3D culture conditions, which reportedly produced 60-65 % functional β islet cells. Here, we describe the protocol for differentiation of human pluripotent stem cells into functional β cells under both 2D and 3D culture conditions.

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Mendeley readers

The data shown below were compiled from readership statistics for 1,070 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
United States 9 <1%
France 4 <1%
Brazil 3 <1%
Japan 3 <1%
Spain 2 <1%
Netherlands 1 <1%
United Kingdom 1 <1%
Ukraine 1 <1%
Iran, Islamic Republic of 1 <1%
Other 5 <1%
Unknown 1040 97%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 273 26%
Researcher 212 20%
Student > Master 175 16%
Student > Bachelor 110 10%
Student > Doctoral Student 62 6%
Other 164 15%
Unknown 74 7%
Readers by discipline Count As %
Agricultural and Biological Sciences 311 29%
Biochemistry, Genetics and Molecular Biology 298 28%
Medicine and Dentistry 101 9%
Engineering 71 7%
Immunology and Microbiology 43 4%
Other 137 13%
Unknown 109 10%