Current Protocols in Stem Cell Biology
Last Update: May 09, 2013
Print ISSN: 1941-7322
Online ISSN: 1938-8969
- Overview
- Table of Contents
- New Protocols
- Sample Unit
- Editors & Contributors
Table of Contents
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Chapter 1 Embryonic and Extraembryonic Stem Cells
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Section A Isolation of Embryonic Stem Cells
- Unit 1A.1 Derivation and Characterization of Nonhuman Primate Embryonic Stem Cells
- Unit 1A.2 Derivation of hESC from Intact Blastocysts
- Unit 1A.3 Reprogramming Primordial Germ Cells (PGC) to Embryonic Germ (EG) Cells
- Unit 1A.4 Derivation and Propagation of hESC Under a Therapeutic Environment
- Unit 1A.5 Generation of Human Embryonic Stem Cells
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Section B Characterization of Embryonic Stem Cells
- Unit 1B.1 Proteomic Analysis of Pluripotent Stem Cells
- Unit 1B.2 Gene Expression Analysis of RNA Purified from Embryonic Stem Cells and Embryoid Body–Derived Cells Using a High‐Throughput Microarray Platform
- Unit 1B.3 Phenotypic Analysis of Human Embryonic Stem Cells
- Unit 1B.4 Isolation of Human Embryonic Stem Cell–Derived Teratomas for the Assessment of Pluripotency
- Unit 1B.5 Tandem Affinity Purification of Protein Complexes in Mouse Embryonic Stem Cells Using In Vivo Biotinylation
- Unit 1B.6 Characterization of X‐Chromosome Inactivation Status in Human Pluripotent Stem Cells
- Unit 1B.7 Preparation of Defined Human Embryonic Stem Cell Populations for Transcriptional Profiling
- Unit 1B.8 Identifying Translationally Regulated Genes During Stem Cell Differentiation
- Unit 1B.9 Quantification of Pluripotency Transcription Factor Levels in Embryonic Stem Cells by Flow Cytometry
- Unit 1B.10 The Streptavidin/Biotinylated DNA/Protein Bound Complex Protocol for Determining the Association of c‐JUN Protein with NANOG Promoter
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Section C Culture and Maintenance of Undifferentiated Embryonic Stem Cells
- Unit 1C.1 Expansion of Human Embryonic Stem Cells In Vitro
- Unit 1C.2 Defined, Feeder‐Independent Medium for Human Embryonic Stem Cell Culture
- Unit 1C.3 Isolation and Propagation of Mouse Embryonic Fibroblasts and Preparation of Mouse Embryonic Feeder Layer Cells
- Unit 1C.4 Culture of Mouse Embryonic Stem Cells
- Unit 1C.5 Preparation of Autogenic Human Feeder Cells for Growth of Human Embryonic Stem Cells
- Unit 1C.6 Isolation of Human Placental Fibroblasts
- Unit 1C.7 Derivation of Human Skin Fibroblast Lines for Feeder Cells of Human Embryonic Stem Cells
- Unit 1C.8 Cryopreservation of Dissociated Human Embryonic Stem Cells in the Presence of ROCK Inhibitor
- Unit 1C.9 Authentication and Banking of Human Pluripotent Stem Cells
- Unit 1C.10 Clump Passaging and Expansion of Human Embryonic and Induced Pluripotent Stem Cells on Mouse Embryonic Fibroblast Feeder Cells
- Unit 1C.11 Expansion of Human Embryonic Stem Cells on Cellulose Microcarriers
- Unit 1C.12 Live‐Cell Immunofluorescence Staining of Human Pluripotent Stem Cells
- Unit 1C.13 Adaptation of Stem Cells to 96‐Well Plate Assays: Use of Human Embryonic and Mouse Neural Stem Cells in the MTT Assay
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Section D Germ Layer Induction/Differentiation of Embryonic Stem Cells
- Unit 1D.1 Germ Layer Induction in ESC—Following the Vertebrate Roadmap
- Unit 1D.2 Formation and Hematopoietic Differentiation of Human Embryoid Bodies by Suspension and Hanging Drop Cultures
- Unit 1D.3 Directed Differentiation of Human Embryonic Stem Cells as Spin Embryoid Bodies and a Description of the Hematopoietic Blast Colony Forming Assay
- Unit 1D.4 Differentiation of Human Embryonic Stem Cells in Adherent and in Chemically Defined Culture Conditions
- Unit 1D.5 Isolation and Differentiation of Xenopus Animal Cap Cells
- Unit 1D.6 High‐Throughput Screening Assay for Embryoid Body Differentiation of Human Embryonic Stem Cells
- Unit 1D.7 Differentiation of Human Embryonic Stem Cells to Cardiomyocytes on Microcarrier Cultures
- Unit 1D.8 Using ES Cells Labeled with GFP for Analyzing Cell Behavior During Differentiation
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Section E Extraembryonic Lineages
- Unit 1E.1 Isolation of Human Placenta‐Derived Multipotent Cells and In Vitro Differentiation into Hepatocyte‐Like Cells
- Unit 1E.2 Isolation of Mesenchymal Stem Cells from Amniotic Fluid and Placenta
- Unit 1E.3 Isolation of Amniotic Epithelial Stem Cells
- Unit 1E.4 Isolation and Manipulation of Mouse Trophoblast Stem Cells
- Unit 1E.5 Isolation of Amniotic Mesenchymal Stem Cells
- Unit 1E.6 Amnion Epithelial Cell Isolation and Characterization for Clinical Use
- Unit 1E.7 A Protocol to Quantify Mammary Early Common Progenitors from Long‐Term Mammosphere Culture
- Unit 1E.8 Amniotic Membrane: Separation of Amniotic Mesoderm from Amniotic Epithelium and Isolation of Their Respective Mesenchymal Stromal and Epithelial Cells
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Section F Mesodermal Lineages
- Unit 1F.1 Differentiation of Embryonic Stem Cells into Cartilage Cells
- Unit 1F.2 Differentiation of Human Embryonic Stem Cells to Cardiomyocytes by Coculture with Endoderm in Serum‐Free Medium
- Unit 1F.3 Isolation of Hematopoietic Stem Cells from Mouse Embryonic Stem Cells
- Unit 1F.4 Differentiation of Mouse Embryonic Stem Cells into Blood
- Unit 1F.5 Endothelial Differentiation of Embryonic Stem Cells
- Unit 1F.6 Hematopoietic Differentiation of Human Embryonic Stem Cells by Cocultivation with Stromal Layers
- Unit 1F.7 TLX1 (HOX11) Immortalization of Embryonic Stem Cell–Derived and Primary Murine Hematopoietic Progenitors
- Unit 1F.8 Differentiation of Multipotent Mesenchymal Precursors and Skeletal Myoblasts from Human Embryonic Stem Cells
- Unit 1F.9 Derivation of Vasculature from Embryonic Stem Cells
- Unit 1F.10 Isolation and Functional Characterization of Pluripotent Stem Cell–Derived Cardiac Progenitor Cells
- Unit 1F.11 Differentiation of Mouse Embryonic Stem Cells into Cardiomyocytes via the Hanging‐Drop and Mass Culture Methods
- Unit 1F.12 Lentiviral Transduction and Clonal Selection of hESCs with Endothelial‐Specific Transgenic Reporters
- Unit 1F.13 Serum‐Free Generation of Multipotent Mesoderm (Kdr+) Progenitor Cells in Mouse Embryonic Stem Cells for Functional Genomics Screening
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Section G Endodermal Lineages
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Section H Ectodermal Lineages
- Unit 1H.1 Differentiation of Mouse Embryonic Stem Cells to Spinal Motor Neurons
- Unit 1H.2 Time‐Lapse Imaging of Embryonic Neural Stem Cell Division in Drosophila by Two‐Photon Microscopy
- Unit 1H.3 Basic Techniques in Human Mesenchymal Stem Cell Cultures: Differentiation into Osteogenic and Adipogenic Lineages, Genetic Perturbations, and Phenotypic Analyses
- Unit 1H.4 Isolation and Culture of Adult Ciliary Epithelial Cells, Previously Identified as Retinal Stem Cells, and Retinal Progenitor Cells
- Unit 1H.5 Derivation of Cerebellar Neurons from Human Pluripotent Stem Cells
- Unit 1H.6 Dopaminergic Differentiation of Human Pluripotent Cells
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Chapter 2 Somatic Stem Cells
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Section A Hematopoietic Stem Cells
- Unit 2A.1 Isolation of Mononuclear Cells from Human Cord Blood by Ficoll‐Paque Density Gradient
- Unit 2A.2 Isolation of Hematopoietic Stem Cells from Human Cord Blood
- Unit 2A.3 Isolation of Mesenchymal Stem Cells from Human Cord Blood
- Unit 2A.4 Isolation and Assessment of Long‐Term Reconstituting Hematopoietic Stem Cells from Adult Mouse Bone Marrow
- Unit 2A.5 Analysis of the Hematopoietic Stem Cell Niche
- Unit 2A.6 Analysis and Manipulation of Hematopoietic Progenitor and Stem Cells from Murine Embryonic Tissues
- Unit 2A.7 High Level In Vitro Expansion of Murine Hematopoietic Stem Cells
- Unit 2A.8 Isolation and Visualization of Mouse Placental Hematopoietic Stem Cells
- Unit 2A.9 Preparation of Hematopoietic Stem and Progenitor Cells from the Human Placenta
- Unit 2A.10 Isolation of Human Umbilical Cord Blood Aldehyde Dehydrogenase–Expressing Progenitor Cells that Modulate Vascular Regenerative Functions In Vitro and In Vivo
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Section B Non‐Hematopoietic Bone Marrow‐Derived Stem Cells
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Section C Cardiovascular Stem Cells
- Unit 2C.1 Isolation and Characterization of Endothelial Progenitor Cells from Human Blood
- Unit 2C.2 Derivation of Epicardium‐Derived Progenitor Cells (EPDCs) from Adult Epicardium
- Unit 2C.3 Isolation and Expansion of Cardiosphere‐Derived Stem Cells
- Unit 2C.4 Expanding Donor Muscle‐Derived Cells for Transplantation
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Section D Neural Stem Cells
- Unit 2D.1 Generation of Cultured Oligodendrocyte Progenitor Cells from Rat Neonatal Brains
- Unit 2D.2 Isolating, Expanding, and Infecting Human and Rodent Fetal Neural Progenitor Cells
- Unit 2D.3 Long‐Term Multilayer Adherent Network (MAN) Expansion, Maintenance, and Characterization, Chemical and Genetic Manipulation, and Transplantation of Human Fetal Forebrain Neural Stem Cells
- Unit 2D.4 Culture System for Rodent and Human Oligodendrocyte Specification, Lineage Progression, and Maturation
- Unit 2D.5 Isolation and Culture of Ventral Mesencephalic Precursor Cells and Dopaminergic Neurons from Rodent Brains
- Unit 2D.6 Isolation of Neural Stem Cells from Neural Tissues Using the Neurosphere Technique
- Unit 2D.7 Neural Stem Cell Grafting in an Animal Model of Chronic Temporal Lobe Epilepsy
- Unit 2D.8 Reproducible Expansion and Characterization of Mouse Neural Stem/Progenitor Cells in Adherent Cultures Derived from the Adult Subventricular Zone
- Unit 2D.9 Neural Differentiation of P19 Carcinoma Cells and Primary Neurospheres: Cell Morphology, Proliferation, Viability, and Functionality
- Unit 2D.10 Single and Dual Birthdating Procedures for Assessing the Response of Adult Neural Stem Cells to the Infusion of a Soluble Factor Using Halogenated Thymidine Analogs
- Unit 2D.11 In Vitro Generation of Three‐Dimensional Substrate‐Adherent Embryonic Stem Cell–Derived Neural Aggregates for Application in Animal Models of Neurological Disorders
- Unit 2D.12 Scalable Production of Transplantable Dopaminergic Neurons from hESCs and iPSCs in Xeno‐Free Defined Conditions
- Unit 2D.13 Functional Stem Cell Integration Assessed by Organotypic Slice Cultures
- Unit 2D.14 Evaluation of Proliferation of Neural Stem Cells In Vitro and In Vivo
- Unit 2D.15 Revealing Neuronal Circuitry Using Stem Cell‐Derived Neurons
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Section E Germline Stem Cells
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Section F Gut Stem Cells
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Section G Lung Stem Cells
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Chapter 3 Cancer Stem Cells
- Unit 3.1 Colon Cancer Stem Cells
- Unit 3.2 In Vivo Evaluation of Leukemic Stem Cells through the Xenotransplantation Model
- Unit 3.3 Culture and Isolation of Brain Tumor Initiating Cells
- Unit 3.4 Isolation and Expansion of Regionally Defined Human Glioblastoma Cells In Vitro
- Unit 3.5 Organotypic Explant Culture of Glioblastoma Multiforme and Subsequent Single‐Cell Suspension
- Unit 3.6 Culture and Isolation of Melanoma‐Initiating Cells
- Unit 3.7 In Vitro Enrichment of Tumor‐Initiating Cells from Human Established Cell Lines
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Chapter 4 Manipulation of Potency
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Section A iPS Cells
- Unit 4A.1 Human iPS Cell Derivation/Reprogramming
- Unit 4A.2 Generation and Characterization of Human Induced Pluripotent Stem Cells
- Unit 4A.3 Derivation of Induced Pluripotent Stem Cells from Human Peripheral Circulating T Cells
- Unit 4A.4 Using MicroRNAs to Enhance the Generation of Induced Pluripotent Stem Cells
- Unit 4A.5 Excision of a Viral Reprogramming Cassette by Delivery of Synthetic Cre mRNA
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Section B Nuclear Transfer
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Chapter 5 Genetic Manipulation of Stem Cells
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Section A Lineage Tracers in Stem Cells
- Unit 5A.1 Imaging Neural Stem Cell Fate in Mouse Model of Glioma
- Unit 5A.2 Functional Analysis of Adult Stem Cells Using Cre‐Mediated Lineage Tracing
- Unit 5A.3 Magnetic Resonance Imaging of Human Embryonic Stem Cells
- Unit 5A.4 Lineage Tracing in the Intestinal Epithelium
- Unit 5A.5 Kaede‐Centrin1 Labeling of Mother and Daughter Centrosomes in Mammalian Neocortical Neural Progenitors
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Section B Homologous Recombination in Stem Cells
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Section C RNA Interference in Stem Cells
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Appendix 1 Useful Information
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Appendix 2 Laboratory Equipment
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