Includes cutting-edge methods and protocols in embryonic stem cell research
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
Includes cutting-edge methods and protocols in embryonic stem cell research
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
This extensive volume explores areas of intense activity related to the very early commitment of stem cells to particular lineages and the progression of differentiation to mature cell stages. Research on embryonic stem cells continues to move very quickly, thus the kinds of studies continue to expand and diversify, and methodologies are continuously being refined and improved, which this book reflects. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions on their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and fully updated, Embryonic Stem Cell Protocols, Third Edition serves as an ideal reference for researchers investigating this rich area of study.
1. Maximizing Clonal Embryonic Stem Cell Derivation by ERK Pathway Inhibition
Jennifer Nichols and Thorsten Boroviak
2. A Simple and Efficient Method of Slow Freezing for Human Embryonic Stem Cells and Induced Pluripotent Stem Cells
Keitaro Imaizumi, Momoe Iha, Naoki Nishishita, Shin Kawamata, Shinichi Nishikawa, and Teruo Akuta
3. Resolving Heterogeneity: Fluorescence Activated Cell Sorting of Dynamic Cell Populations from Feeder-Free Mouse Embryonic Stem Cell Culture
Jurriaan J. Hölzenspies, Gelo V. Dela Cruz, and Joshua M. Brickman
4. Acquiring Ground State Pluripotency: Switching Mouse Embryonic Stem Cells from Serum/LIF Medium to 2i/LIF Medium
Matteo Tosolini and Alice Jouneau
5. Generation of Embryonic Stem Cells in Rabbits
Marielle Afanassieff, Pierre Osteil, and Pierre Savatier
6. Gene Transfer into Pluripotent Stem Cells Via Lentiviral Transduction
Ortwin Naujok, Ulf Diekmann, and Matthias Elsner
7. Imaging Pluripotency: Time-Lapse Analysis of Mouse Embryonic Stem Cells
Anna Pezzarossa, Ana M.V. Guedes, Domingos Henrique, and Elsa Abranches
8. Expanding the Utility of FUCCI Reporters Using FACS-Based `Omics Analysis
James Chappell, Ben Boward, and Stephen Dalton
9. Selection of Antibodies Interfering with Cell-Surface Receptor Signalling Using Embryonic Stem Cell Differentiation
Anna N. Melidoni, Michael R. Dyson, and John McCafferty
10. Delivering Antisense Morpholino Oligonucleotides to Target Telomerase Splice Variants in Human Embryonic Stem Cells
Lida Radan, Chris S. Hughes, Jonathan H. Teichroeb, Lynne-Marie Postovit, and Dean H. Betts
11. Analysis of mRNA Translation Rate in Mouse Embryonic Stem Cells
Anisa B. Rahim and Leah A. Vardy
12. Generation and Purification of Definitive Endoderm Cells Generated from Pluripotent Stem Cells
Ulf Diekmann and Ortwin Naujok
13. Definitive Endoderm Differentiation of Human Embryonic Stem Cells Combined with Selective Elimination of Undifferentiated Cells by Methionine Deprivation
Tomonori Tsuyama, Nobuaki Shiraki, and Shoen Kume
14. Selective Differentiation into Hematopoietic and Cardiac Cells from Pluripotent Stem Cells Based on the Expression of Cell Surface Markers
Atsumasa Okada, Katsuhisa Tashiro, Tomoko Yamaguchi, and Kenji Kawabata
15. Methods for Derivation of Multipotent Neural Crest Cells Derived from Human Pluripotent Stem Cells
John Avery and Stephen Dalton
16. From Naive to Primed Pluripotency: In Vitro Conversion of Mouse Embryonic Stem Cells in Epiblast Stem Cells
Matteo Tosolini and Alice Jouneau
17. In Vitro Differentiation of Embryonic Stem Cells into Hematopoietic Lineage: Towards Erythroid Progenitor´s Production
Iliana Fauzi, Nicki Panoskaltsis, and Athanasios Mantalaris
18. Dopaminergic Differentiation of Human Embryonic Stem Cells on PA6-Derived Adipocytes
M. Oktar Guloglu and Anna Larsen
19. Intracellular Calcium Measurements for Functional Characterization of Neuronal Phenotypes
Talita Glaser, Ana Regina G. Castillo, Ágatha Oliveira, and Henning Ulrich
20. In Vitro Differentiation of Pluripotent Stem Cells into Functional ? Islets Under 2D and 3D Culture Conditions and In Vivo Preclinical Validation of 3D Islets
Bipasha Bose and Sudheer Shenoy P
21. Generation of Corneal Keratocytes from Human Embryonic Stem Cells
Andrew J. Hertsenberg and James L. Funderburgh
22. Maintenance, Transgene Delivery, and Pluripotency Measurement of Mouse Embryonic Stem Cells
Tetsuya S. Tanaka
23. Generation of a Knockout Mouse Embryonic Stem Cell Line Using a Paired CRISPR/Cas9 Genome Engineering Tool
Rahel Wettstein, Maxime Bodak, and Constance Ciaudo
24. Derivation of Neural Precursor Cells from Human Embryonic Stem Cells for DNA Methylomic Analysis
Ivan Roubal, Sun Joo Park, and Yong Kim
25. 26S and PA28-20S Proteasome Activity in Cytosolic Extracts from Embryonic Stem Cells
Malin Hernebring
26. A Concise Protocol for siRNA-Mediated Gene Suppression in Human Embryonic Stem Cells
Peter F. Renz and Tobias A. Beyer
27. Applying Shear Stress to Pluripotent Stem Cells
Russell P. Wolfe, Julia B. Guidry, Stephanie L. Messina, and Tabassum Ahsan
28. Generating Inner Ear Organoids from Mouse Embryonic Stem Cells
Emma Longworth-Mills, Karl R. Koehler, and Eri Hashino
29. Differentiation of Adipocytes in Monolayer from Mouse Embryonic Stem Cells
Ixchelt Cuaranta-Monroy, Zoltan Simandi, and Laszlo Nagy
30. Pancreatic Differentiation from Murine Embryonic Stem Cells
Daisuke Sakano, Nobuaki Shiraki, and Shoen Kume
31. Osteogenic Differentiation from Embryonic Stem Cells
Yanhong Yu, Carlos Pilquil, and Michal Opas
32. Limbal Stromal Tissue Specific Stem Cells and Their Differentiation Potential to Corneal Epithelial Cells
Kishore Reddy Katikireddy and Ula V. Jurkunas
This extensive volume explores areas of intense activity related to the very early commitment of stem cells to particular lineages and the progression of differentiation to mature cell stages. Research on embryonic stem cells continues to move very quickly, thus the kinds of studies continue to expand and diversify, and methodologies are continuously being refined and improved, which this book reflects. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions on their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and fully updated,
Embryonic Stem Cell Protocols, Third Edition serves as an ideal reference for researchers investigating this rich area of study.
Maximizing Clonal Embryonic Stem Cell Derivation by ERK Pathway Inhibition.- A Simple and Efficient Method of Slow Freezing for Human Embryonic Stem Cells and Induced Pluripotent Stem Cells.- Resolving Heterogeneity: Fluorescence Activated Cell Sorting of Dynamic Cell Populations from Feeder-Free Mouse Embryonic Stem Cell Culture.- Acquiring Ground State Pluripotency: Switching Mouse Embryonic Stem Cells from Serum/LIF Medium to 2i/LIF Medium.- Generation of Embryonic Stem Cells in Rabbits.- Gene Transfer into Pluripotent Stem Cells Via Lentiviral Transduction.- Imaging Pluripotency: Time-Lapse Analysis of Mouse Embryonic Stem Cells.- Expanding the Utility of FUCCI Reporters Using FACS-Based 'Omics Analysis.- Selection of Antibodies Interfering with Cell-Surface Receptor Signalling Using Embryonic Stem Cell Differentiation.- Delivering Antisense Morpholino Oligonucleotides to Target Telomerase Splice Variants in Human Embryonic Stem Cells.- Analysis of mRNA Translation Rate in Mouse Embryonic Stem Cells.- Generation and Purification of Definitive Endoderm Cells Generated from Pluripotent Stem Cells.- Definitive Endoderm Differentiation of Human Embryonic Stem Cells Combined with Selective Elimination of Undifferentiated Cells by Methionine Deprivation.- Selective Differentiation into Hematopoietic and Cardiac Cells from Pluripotent Stem Cells Based on the Expression of Cell Surface Markers.- Methods for Derivation of Multipotent Neural Crest Cells Derived from Human Pluripotent Stem Cells.- From Naive to Primed Pluripotency: In Vitro Conversion of Mouse Embryonic Stem Cells in Epiblast Stem Cells.- In Vitro Differentiation of Embryonic Stem Cells into Hematopoietic Lineage: Towards Erythroid Progenitor's Production.- Dopaminergic Differentiation of Human Embryonic Stem Cells on PA6-Derived Adipocytes.- Intracellular Calcium Measurements for Functional Characterization ofNeuronal Phenotypes.- In Vitro Differentiation of Pluripotent Stem Cells into Functional beta Islets Under 2D and 3D Culture Conditions and In Vivo Preclinical Validation of 3D Islets.- Generation of Corneal Keratocytes from Human Embryonic Stem Cells.- Maintenance, Transgene Delivery, and Pluripotency Measurement of Mouse Embryonic Stem Cells.- Generation of a Knockout Mouse Embryonic Stem Cell Line Using a Paired CRISPR/Cas9 Genome Engineering Tool.- Derivation of Neural Precursor Cells from Human Embryonic Stem Cells for DNA Methylomic Analysis.- 26S and PA28-20S Proteasome Activity in Cytosolic Extracts from Embryonic Stem Cells.- A Concise Protocol for siRNA-Mediated Gene Suppression in Human Embryonic Stem Cells.- Applying Shear Stress to Pluripotent Stem Cells.- Generating Inner Ear Organoids from Mouse Embryonic Stem Cells.- Differentiation of Adipocytes in Monolayer from Mouse Embryonic Stem Cells.- Pancreatic Differentiation from Murine Embryonic Stem Cells.- Osteogenic Differentiation from Embryonic Stem Cells.- Limbal Stromal Tissue Specific Stem Cells and Their Differentiation Potential to Corneal Epithelial Cells.
Inhaltsverzeichnis
1. Maximizing Clonal Embryonic Stem Cell Derivation by ERK Pathway Inhibition
Jennifer Nichols and Thorsten Boroviak
2. A Simple and Efficient Method of Slow Freezing for Human Embryonic Stem Cells and Induced Pluripotent Stem Cells
Keitaro Imaizumi, Momoe Iha, Naoki Nishishita, Shin Kawamata, Shinichi Nishikawa, and Teruo Akuta
3. Resolving Heterogeneity: Fluorescence Activated Cell Sorting of Dynamic Cell Populations from Feeder-Free Mouse Embryonic Stem Cell Culture
Jurriaan J. Hölzenspies, Gelo V. Dela Cruz, and Joshua M. Brickman
4. Acquiring Ground State Pluripotency: Switching Mouse Embryonic Stem Cells from Serum/LIF Medium to 2i/LIF Medium
Matteo Tosolini and Alice Jouneau
5. Generation of Embryonic Stem Cells in Rabbits
Marielle Afanassieff, Pierre Osteil, and Pierre Savatier
6. Gene Transfer into Pluripotent Stem Cells Via Lentiviral Transduction
Ortwin Naujok, Ulf Diekmann, and Matthias Elsner
7. Imaging Pluripotency: Time-Lapse Analysis of Mouse Embryonic Stem Cells
Anna Pezzarossa, Ana M.V. Guedes, Domingos Henrique, and Elsa Abranches
8. Expanding the Utility of FUCCI Reporters Using FACS-Based `Omics Analysis
James Chappell, Ben Boward, and Stephen Dalton
9. Selection of Antibodies Interfering with Cell-Surface Receptor Signalling Using Embryonic Stem Cell Differentiation
Anna N. Melidoni, Michael R. Dyson, and John McCafferty
10. Delivering Antisense Morpholino Oligonucleotides to Target Telomerase Splice Variants in Human Embryonic Stem Cells
Lida Radan, Chris S. Hughes, Jonathan H. Teichroeb, Lynne-Marie Postovit, and Dean H. Betts
11. Analysis of mRNA Translation Rate in Mouse Embryonic Stem Cells
Anisa B. Rahim and Leah A. Vardy
12. Generation and Purification of Definitive Endoderm Cells Generated from Pluripotent Stem Cells
Ulf Diekmann and Ortwin Naujok
13. Definitive Endoderm Differentiation of Human Embryonic Stem Cells Combined with Selective Elimination of Undifferentiated Cells by Methionine Deprivation
Tomonori Tsuyama, Nobuaki Shiraki, and Shoen Kume
14. Selective Differentiation into Hematopoietic and Cardiac Cells from Pluripotent Stem Cells Based on the Expression of Cell Surface Markers
Atsumasa Okada, Katsuhisa Tashiro, Tomoko Yamaguchi, and Kenji Kawabata
15. Methods for Derivation of Multipotent Neural Crest Cells Derived from Human Pluripotent Stem Cells
John Avery and Stephen Dalton
16. From Naive to Primed Pluripotency: In Vitro Conversion of Mouse Embryonic Stem Cells in Epiblast Stem Cells
Matteo Tosolini and Alice Jouneau
17. In Vitro Differentiation of Embryonic Stem Cells into Hematopoietic Lineage: Towards Erythroid Progenitor¿s Production
Iliana Fauzi, Nicki Panoskaltsis, and Athanasios Mantalaris
18. Dopaminergic Differentiation of Human Embryonic Stem Cells on PA6-Derived Adipocytes
M. Oktar Guloglu and Anna Larsen
19. Intracellular Calcium Measurements for Functional Characterization of Neuronal Phenotypes
Talita Glaser, Ana Regina G. Castillo, Ágatha Oliveira, and Henning Ulrich
20. In Vitro Differentiation of Pluripotent Stem Cells into Functional ß Islets Under 2D and 3D Culture Conditions and In Vivo Preclinical Validation of 3D Islets
Bipasha Bose and Sudheer Shenoy P
21. Generation of Corneal Keratocytes from Human Embryonic Stem Cells
Andrew J. Hertsenberg and James L. Funderburgh
22. Maintenance, Transgene Delivery, and Pluripotency Measurement of Mouse Embryonic Stem Cells
Tetsuya S. Tanaka
23. Generation of a Knockout Mouse Embryonic Stem Cell Line Using a Paired CRISPR/Cas9 Genome Engineering Tool
Rahel Wettstein, Maxime Bodak, and Constance Ciaudo
24. Derivation of Neural Precursor Cells from Human Embryonic Stem Cells for DNA Methylomic Analysis
Ivan Roubal, Sun Joo Park, and Yong Kim
25. 26S and PA28-20S Proteasome Activity in Cytosolic Extracts from Embryonic Stem Cells
Malin Hernebring
26. A Concise Protocol for siRNA-Mediated Gene Suppression in Human Embryonic Stem Cells
Peter F. Renz and Tobias A. Beyer
27. Applying Shear Stress to Pluripotent Stem Cells
Russell P. Wolfe, Julia B. Guidry, Stephanie L. Messina, and Tabassum Ahsan
28. Generating Inner Ear Organoids from Mouse Embryonic Stem Cells
Emma Longworth-Mills, Karl R. Koehler, and Eri Hashino
29. Differentiation of Adipocytes in Monolayer from Mouse Embryonic Stem Cells
Ixchelt Cuaranta-Monroy, Zoltan Simandi, and Laszlo Nagy
30. Pancreatic Differentiation from Murine Embryonic Stem Cells
Daisuke Sakano, Nobuaki Shiraki, and Shoen Kume
31. Osteogenic Differentiation from Embryonic Stem Cells
Yanhong Yu, Carlos Pilquil, and Michal Opas
32. Limbal Stromal Tissue Specific Stem Cells and Their Differentiation Potential to Corneal Epithelial Cells
Kishore Reddy Katikireddy and Ula V. Jurkunas
Klappentext
This extensive volume explores areas of intense activity related to the very early commitment of stem cells to particular lineages and the progression of differentiation to mature cell stages. Research on embryonic stem cells continues to move very quickly, thus the kinds of studies continue to expand and diversify, and methodologies are continuously being refined and improved, which this book reflects. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions on their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and fully updated, Embryonic Stem Cell Protocols, Third Edition serves as an ideal reference for researchers investigating this rich area of study.
Includes cutting-edge methods and protocols in embryonic stem cell research
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
