The purpose of Stem Cell Culture is to provide a comprehensive resource for researchers in the fields of embyonic, fetal and adult stem cell biology to find methods for the purification, culture, and differentitation of these cell types, with the main emphasis on the maintenance of the stem cell phenotype in vitro. This volume will be the first to broadly cover multiple types of stem cell culture from different ages, organs and species. Authors will focus on the practical do's and don'ts of isolating and culturing these cell types, and feel free to use illustrative data or diagrams whereever…mehr
The purpose of Stem Cell Culture is to provide a comprehensive resource for researchers in the fields of embyonic, fetal and adult stem cell biology to find methods for the purification, culture, and differentitation of these cell types, with the main emphasis on the maintenance of the stem cell phenotype in vitro. This volume will be the first to broadly cover multiple types of stem cell culture from different ages, organs and species. Authors will focus on the practical do's and don'ts of isolating and culturing these cell types, and feel free to use illustrative data or diagrams whereever this improves the comprehension of the reader. This should allow the reader to compare and contrast techniques and make this a standard reference for those in the field, or desiring to start stem cell culture.
Derivation of Human Embryonic Stem Cells in Standard and Chemically Defined Conditions
I Introduction
II Methods
III Discussion
IV Materials
Autogeneic Feeders for the Culture of Undifferentiated Human Embryonic Stem Cells in Feeder and Feederâ?Free Conditions
I Introduction
II Methods and Materials
III Results
IV Discussion and Summary
Microporous Membrane Growth Substrates for Embryonic Stem Cell Culture and Differentiation
I Introduction
II Rationale
III Methods
IV Summary
Culture of Rodent Spermatogonial Stem Cells, Male Germline Stem Cells of the Postnatal Animal
I Introduction
II Rationale
III Methods
IV Materials
V Discussion
Characterization of Human Amniotic Fluid Stem Cells and Their Pluripotential Capability
I Introduction
II Amniotic Fluid Cell Composition and Stem Cells
III In Vitro Potential of Amniotic Fluid Stem Cells
IV Ex Vivo and In Vivo Application of Amniotic Fluid Stem Cells
V Discussion
Method to Isolate Mesenchymalâ?Like Cells from Wharton's Jelly of Umbilical Cord
I Introduction
II Rationale
III Methods
IV Materials
V UCMSCs and Culture Characteristics
VI Discussion
Isolation, Characterization, and Differentiation of Human Umbilical Cord Perivascular Cells (HUCPVCs)
I Introduction
II Methods
III Concluding Remarks
Hepatic Stem Cells and Hepatoblasts: Identification, Isolation, and Ex Vivo Maintenance
I Introduction
II Epithelialâ?"Mesenchymal Relationship
III Stem Cells and Lineage Biology
IV Liver Processing
V Fractionation of Liver Cell Subpopulations
VI Feeder Cells
VII Extracellular Matrix
VIII Culture Media
IX Monolayer Cultures of Cells
X Threeâ?Dimensional Systems
Acknowledgments
Appendix 1: Nomenclature, Glossary, and Abbreviations
Appendix 2: Details on the Preparation of Buffers/Reagents
Appendix 3: Commercial Sources of Reagents
Appendix 4: Sources of Primary and Secondary Antibodies for Immunoselection and for Characterization of the Cells. (See Below for Suppliers and the Location of the Suppliers)
Culture of Pluripotent Neural Epithelial Progenitor Cells from E9 Rat Embryo
I Introduction
II Rationale
III Methods
IV Materials
V Discussion
VI Summary
Primary and Multipassage Culture of Human Fetal Kidney Epithelial Progenitor Cells
I Introduction
II Rationale
III Methods
IV Materials
V Discussion
VI Summary
Isolation of Human Mesenchymal Stem Cells from Bone and Adipose Tissue
I Introduction
II Isolation of hMSCs from Trabecular Bone
III Isolation of hMSCs from Adipose Tissue
IV Growth, Subculture, and Cryopreservation of Undifferentiated hMSCs
V Surface Marker Characterization of hMSCs
VI hMSC Differentiation Assays for Osteogenic, Adipogenic, and Chondrogenic Pathways
VII Summary
Culture of Mesenchymal Stem/Progenitor Cells in Adhesionâ?Independent Conditions
I Introduction
II Suspension Culture of Bone Marrowâ?Derived MPCs
III Analysis of Suspension Culture
IV Mechanism of MPC Expansion in Suspension
V Discussion
Purification and Longâ?Term Culture of Multipotent Progenitor Cells Affiliated with the Walls of Human Blood Vessels: Myoendothelial Cells and Pericytes
I Introduction
II Materials and Methods: Cell Analysis and Purification by Flow Cytometry
III Materials and Methods: Longâ?Term Culture of Human Blood Vesselâ?Associated Progenitor Cells
IV Materials and Methods: Proliferation Kinetics of Human Myoendothelial Cell
Derivation of Human Embryonic Stem Cells in Standard and Chemically Defined Conditions
I Introduction
II Methods
III Discussion
IV Materials
Autogeneic Feeders for the Culture of Undifferentiated Human Embryonic Stem Cells in Feeder and Feederâ?Free Conditions
I Introduction
II Methods and Materials
III Results
IV Discussion and Summary
Microporous Membrane Growth Substrates for Embryonic Stem Cell Culture and Differentiation
I Introduction
II Rationale
III Methods
IV Summary
Culture of Rodent Spermatogonial Stem Cells, Male Germline Stem Cells of the Postnatal Animal
I Introduction
II Rationale
III Methods
IV Materials
V Discussion
Characterization of Human Amniotic Fluid Stem Cells and Their Pluripotential Capability
I Introduction
II Amniotic Fluid Cell Composition and Stem Cells
III In Vitro Potential of Amniotic Fluid Stem Cells
IV Ex Vivo and In Vivo Application of Amniotic Fluid Stem Cells
V Discussion
Method to Isolate Mesenchymalâ?Like Cells from Wharton's Jelly of Umbilical Cord
I Introduction
II Rationale
III Methods
IV Materials
V UCMSCs and Culture Characteristics
VI Discussion
Isolation, Characterization, and Differentiation of Human Umbilical Cord Perivascular Cells (HUCPVCs)
I Introduction
II Methods
III Concluding Remarks
Hepatic Stem Cells and Hepatoblasts: Identification, Isolation, and Ex Vivo Maintenance
I Introduction
II Epithelialâ?"Mesenchymal Relationship
III Stem Cells and Lineage Biology
IV Liver Processing
V Fractionation of Liver Cell Subpopulations
VI Feeder Cells
VII Extracellular Matrix
VIII Culture Media
IX Monolayer Cultures of Cells
X Threeâ?Dimensional Systems
Acknowledgments
Appendix 1: Nomenclature, Glossary, and Abbreviations
Appendix 2: Details on the Preparation of Buffers/Reagents
Appendix 3: Commercial Sources of Reagents
Appendix 4: Sources of Primary and Secondary Antibodies for Immunoselection and for Characterization of the Cells. (See Below for Suppliers and the Location of the Suppliers)
Culture of Pluripotent Neural Epithelial Progenitor Cells from E9 Rat Embryo
I Introduction
II Rationale
III Methods
IV Materials
V Discussion
VI Summary
Primary and Multipassage Culture of Human Fetal Kidney Epithelial Progenitor Cells
I Introduction
II Rationale
III Methods
IV Materials
V Discussion
VI Summary
Isolation of Human Mesenchymal Stem Cells from Bone and Adipose Tissue
I Introduction
II Isolation of hMSCs from Trabecular Bone
III Isolation of hMSCs from Adipose Tissue
IV Growth, Subculture, and Cryopreservation of Undifferentiated hMSCs
V Surface Marker Characterization of hMSCs
VI hMSC Differentiation Assays for Osteogenic, Adipogenic, and Chondrogenic Pathways
VII Summary
Culture of Mesenchymal Stem/Progenitor Cells in Adhesionâ?Independent Conditions
I Introduction
II Suspension Culture of Bone Marrowâ?Derived MPCs
III Analysis of Suspension Culture
IV Mechanism of MPC Expansion in Suspension
V Discussion
Purification and Longâ?Term Culture of Multipotent Progenitor Cells Affiliated with the Walls of Human Blood Vessels: Myoendothelial Cells and Pericytes
I Introduction
II Materials and Methods: Cell Analysis and Purification by Flow Cytometry
III Materials and Methods: Longâ?Term Culture of Human Blood Vesselâ?Associated Progenitor Cells
IV Materials and Methods: Proliferation Kinetics of Human Myoendothelial Cell
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