This detailed volume provides thorough protocols describing how to use genetics to study mouse and zebrafish erythropoiesis in whole animal models and for genetically manipulating cultured mouse and human erythroid cells. Protocols include strategies to analyze circulating red blood cell parameters, as well as progenitor cell status, stage of differentiation, and enucleation. Methods for the study of erythroid cell gene regulation using chromatin immunoprecipitation and chromatin conformation capture are detailed. Furthermore, protocols are provided to examine stress erythropoiesis, erythroblastic islands, and erythroblast structure. The collection also includes reviews on the available mouse models of erythropoiesis, as well as on future considerations for good manufacturing practice in order to translate the manufacture of erythrocytes to the clinic. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Erythropoiesis: Methods and Protocols provides a definitive source for molecular and cellular biology protocols to study erythropoiesis and to move this vital field forward.
|Includes cutting-edge methods and protocols for the study of erythropoiesis
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
1. An Introduction to Erythropoiesis Approaches
Joyce A. Lloyd
2. Using the Zebrafish as an Approach to Examine the Mechanisms of Vertebrate Erythropoiesis
Martin D. Kafina and Barry H. Paw
3. Mouse Models of Erythropoiesis and Associated Diseases
Matthew P. Parker and Kenneth R. Peterson
4. Dissecting Regulatory Mechanisms Using Mouse Fetal Liver-Derived Erythroid Cells Skye C. McIver, Kyle J. Hewitt, Xin Gao, Charu Mehta, Jing Zhang, and Emery H. Bresnick
5. Stress Erythropoiesis Model Systems
&
nbsp; Laura F. Bennett, Chang Liao, and Robert F. Paulson
6. Approaches for Analysis of Erythroid Cell Parameters and Hemoglobinopathies in Mouse Models
Marie Trudel and Josepha-Clara Sedzro
7. Functional Analysis of Erythroid Progenitors by Colony-Forming Assays
James Palis and Anne Koniski
8. Analyzing the Formation, Morphology, and Integrity of Erythroblastic Islands
Jia Hao Yeo, Matthew P. Cosgriff, and Stuart T. Fraser
9. Flow Cytometry (FCM) Analysis and Fluorescence-Activated Cell Sorting (FACS) of Erythroid Cells
Xiuli An and Lixiang Chen
10. Analysis of Erythropoiesis Using Imaging Flow Cytometry
Theodosia Kalfa and Kathleen E. McGrath
11. Flow Cytometric Analysis of Erythroblast Enucleation
Xiuli An and Lixiang Chen
12. High-Resolution Fluorescence Microscope Imaging of Erythroblast Structure
Alyson Smith, Roberta B. Nowak, and Velia M. Fowler
13. Chromatin Immunoprecipitation (ChIP) with Erythroid Samples
Ivan Krivega and Ann Dean
14. Chromosome Conformation Capture (3C and Higher) with Erythroid Samples
Ivan Krivega and
Ann Dean
15. Genome Editing of Erythroid Cell Culture Model Systems
Jinfen J. Yik, Merlin Crossley, and Kate G.R. Quinlan
16. In Vitro Erythroid Differentiation and Lentiviral Knockdown in Human CD34+ Cells from Umbilical Cord Blood
Anna Kovilakath, Safa Mohamad, Farrah Hermes, Shou Zhen Wang, Gordon D. Ginder, and Joyce A. Lloyd
17. Growing and Genetically Manipulating Human Umbilical Cord Blood-Derived Erythroid Progenitor (HUDEP) Cell Lines
Divya S. Vinjamur and Daniel E. Bauer
18. Good Manufacturing Practice (GMP) Translation of Advanced Cellular Therapeutics: Lessons for the Manufacture of Erythrocytes as Medicinal Products
&nbs
p; Neil W.A. McGowan, John D.M. Campbell, and Joanne C. Mountford
This detailed volume provides thorough protocols describing how to use genetics to study mouse and zebrafish erythropoiesis in whole animal models and for genetically manipulating cultured mouse and human erythroid cells. Protocols include strategies to analyze circulating red blood cell parameters, as well as progenitor cell status, stage of differentiation, and enucleation. Methods for the study of erythroid cell gene regulation using chromatin immunoprecipitation and chromatin conformation capture are detailed. Furthermore, protocols are provided to examine stress erythropoiesis, erythroblastic islands, and erythroblast structure. The collection also includes reviews on the available mouse models of erythropoiesis, as well as on future considerations for good manufacturing practice in order to translate the manufacture of erythrocytes to the clinic. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Erythropoiesis: Methods and Protocols provides a definitive source for molecular and cellular biology protocols to study erythropoiesis and to move this vital field forward.
This detailed volume provides thorough protocols describing how to use genetics to study mouse and zebrafish erythropoiesis in whole animal models and for genetically manipulating cultured mouse and human erythroid cells. Protocols include strategies to analyze circulating red blood cell parameters, as well as progenitor cell status, stage of differentiation, and enucleation. Methods for the study of erythroid cell gene regulation using chromatin immunoprecipitation and chromatin conformation capture are detailed. Furthermore, protocols are provided to examine stress erythropoiesis, erythroblastic islands, and erythroblast structure. The collection also includes reviews on the available mouse models of erythropoiesis, as well as on future considerations for good manufacturing practice in order to translate the manufacture of erythrocytes to the clinic. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical,
Erythropoiesis: Methods and Protocols provides a definitive source for molecular and cellular biology protocols to study erythropoiesis and to move this vital field forward.
An Introduction to Erythropoiesis Approaches.- Using the Zebrafish as an Approach to Examine the Mechanisms of Vertebrate Erythropoiesis.- Mouse Models of Erythropoiesis and Associated Diseases.- Dissecting Regulatory Mechanisms Using Mouse Fetal Liver-Derived Erythroid Cells.- Stress Erythropoiesis Model Systems.- Approaches for Analysis of Erythroid Cell Parameters and Hemoglobinopathies in Mouse Models.- Functional Analysis of Erythroid Progenitors by Colony-Forming Assays.- Analyzing the Formation, Morphology, and Integrity of Erythroblastic Islands.- Flow Cytometry (FCM) Analysis and Fluorescence-Activated Cell Sorting (FACS) of Erythroid Cells.- Analysis of Erythropoiesis Using Imaging Flow Cytometry.- Flow Cytometric Analysis of Erythroblast Enucleation.- High-Resolution Fluorescence Microscope Imaging of Erythroblast Structure.- Chromatin Immunoprecipitation (ChIP) with Erythroid Samples.- Chromosome Conformation Capture (3C and Higher) with Erythroid Samples.- Genome Editing of Erythroid Cell Culture Model Systems.- In Vitro Erythroid Differentiation and Lentiviral Knockdown in Human CD34+ Cells from Umbilical Cord Blood.- Growing and Genetically Manipulating Human Umbilical Cord Blood-Derived Erythroid Progenitor (HUDEP) Cell Lines.- Good Manufacturing Practice (GMP) Translation of Advanced Cellular Therapeutics: Lessons for the Manufacture of Erythrocytes as Medicinal Products.
Über den Autor
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Inhaltsverzeichnis
1. An Introduction to Erythropoiesis Approaches
Joyce A. Lloyd
2. Using the Zebrafish as an Approach to Examine the Mechanisms of Vertebrate Erythropoiesis
Martin D. Kafina and Barry H. Paw
3. Mouse Models of Erythropoiesis and Associated Diseases
Matthew P. Parker and Kenneth R. Peterson
4. Dissecting Regulatory Mechanisms Using Mouse Fetal Liver-Derived Erythroid Cells
Skye C. McIver, Kyle J. Hewitt, Xin Gao, Charu Mehta, Jing Zhang, and Emery H. Bresnick
5. Stress Erythropoiesis Model Systems
&
nbsp; Laura F. Bennett, Chang Liao, and Robert F. Paulson
6. Approaches for Analysis of Erythroid Cell Parameters and Hemoglobinopathies in Mouse Models
Marie Trudel and Josepha-Clara Sedzro
7. Functional Analysis of Erythroid Progenitors by Colony-Forming Assays
James Palis and Anne Koniski
8.
Analyzing the Formation, Morphology, and Integrity of Erythroblastic Islands
Jia Hao Yeo, Matthew P. Cosgriff, and Stuart T. Fraser
9. Flow Cytometry (FCM) Analysis and Fluorescence-Activated Cell Sorting (FACS) of Erythroid Cells
Xiuli An and Lixiang Chen
10. Analysis of Erythropoiesis Using Imaging Flow Cytometry
Theodosia Kalfa and Kathleen E. McGrath
11. Flow Cytometric Analysis of Erythroblast Enucleation
Xiuli An and Lixiang Chen
12. High-Resolution Fluorescence Microscope Imaging of Erythroblast Structure
Alyson Smith, Roberta B. Nowak, and Velia M. Fowler
13. Chromatin Immunoprecipitation (ChIP) with Erythroid Samples
Ivan Krivega and Ann Dean
14. Chromosome Conformation Capture (3C and Higher) with Erythroid Samples
Ivan Krivega and
Ann Dean
15. Genome Editing of Erythroid Cell Culture Model Systems
Jinfen J. Yik, Merlin Crossley, and Kate G.R. Quinlan
16. In Vitro Erythroid Differentiation and Lentiviral Knockdown in Human CD34+ Cells from Umbilical Cord Blood
Anna Kovilakath, Safa Mohamad, Farrah Hermes, Shou Zhen Wang, Gordon D. Ginder, and Joyce A. Lloyd
17. Growing and Genetically Manipulating Human Umbilical Cord Blood-Derived Erythroid Progenitor (HUDEP) Cell Lines
Divya S. Vinjamur and Daniel E. Bauer
18. Good Manufacturing Practice (GMP) Translation of Advanced Cellular Therapeutics: Lessons for the Manufacture of Erythrocytes as Medicinal Products
&nbs
p; Neil W.A. McGowan, John D.M. Campbell, and Joanne C. Mountford
Klappentext
This detailed volume provides thorough protocols describing how to use genetics to study mouse and zebrafish erythropoiesis in whole animal models and for genetically manipulating cultured mouse and human erythroid cells. Protocols include strategies to analyze circulating red blood cell parameters, as well as progenitor cell status, stage of differentiation, and enucleation. Methods for the study of erythroid cell gene regulation using chromatin immunoprecipitation and chromatin conformation capture are detailed. Furthermore, protocols are provided to examine stress erythropoiesis, erythroblastic islands, and erythroblast structure. The collection also includes reviews on the available mouse models of erythropoiesis, as well as on future considerations for good manufacturing practice in order to translate the manufacture of erythrocytes to the clinic. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
n
Authoritative and practical, Erythropoiesis: Methods and Protocols provides a definitive source for molecular and cellular biology protocols to study erythropoiesis and to move this vital field forward.
Includes cutting-edge methods and protocols for the study of erythropoiesisProvides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
