Stem Cell Biology

  1. Methodologies in Stem Cell Research
    1. Isolation and Purification
      1. Fluorescence-Activated Cell Sorting (FACS)
        1. Principles and Applications
          1. Marker Selection
            1. Multi-parameter Analysis
              1. Cell Viability Considerations
              2. Magnetic-Activated Cell Sorting (MACS)
                1. Principles and Applications
                  1. Advantages and Limitations
                    1. Positive and Negative Selection
                    2. Density Gradient Centrifugation
                      1. Protocols and Uses
                        1. Ficoll-Paque Separation
                          1. Percoll Gradients
                          2. Other Separation Methods
                            1. Adherence-Based Selection
                              1. Size-Based Separation
                                1. Microfluidic Approaches
                              2. Culture and Expansion
                                1. In Vitro Culture Systems
                                  1. Media Composition
                                    1. Basal Media
                                      1. Serum Requirements
                                        1. Growth Factors
                                          1. Small Molecules
                                          2. Environmental Conditions
                                            1. Temperature
                                              1. CO2 Concentration
                                                1. Oxygen Levels
                                              2. Feeder Layers vs. Feeder-Free Culture
                                                1. Types of Feeder Cells
                                                  1. Mouse Embryonic Fibroblasts
                                                    1. Human Feeder Cells
                                                    2. Defined Substrates
                                                      1. Matrigel
                                                        1. Synthetic Matrices
                                                          1. Recombinant Proteins
                                                        2. Suspension vs. Adherent Culture
                                                          1. Advantages and Disadvantages
                                                            1. Embryoid Body Formation
                                                              1. Spinner Flask Culture
                                                              2. 3D Culture and Organoids
                                                                1. Organoid Formation
                                                                  1. Self-Organization Principles
                                                                    1. Growth Factor Gradients
                                                                    2. Applications in Modeling Development and Disease
                                                                      1. Organ-on-Chip Technologies
                                                                      2. Cryopreservation
                                                                        1. Freezing Protocols
                                                                          1. Cryoprotectants
                                                                            1. Vitrification Methods
                                                                          2. Characterization Techniques
                                                                            1. Immunocytochemistry and Immunohistochemistry
                                                                              1. Antibody Selection
                                                                                1. Detection of Stem Cell Markers
                                                                                  1. Fixation and Permeabilization
                                                                                    1. Signal Amplification
                                                                                    2. Flow Cytometry
                                                                                      1. Surface Marker Analysis
                                                                                        1. Cell Sorting Applications
                                                                                          1. Intracellular Staining
                                                                                            1. Cell Cycle Analysis
                                                                                            2. Gene Expression Analysis
                                                                                              1. Quantitative PCR (qPCR)
                                                                                                1. Primer Design
                                                                                                  1. Reference Genes
                                                                                                    1. Data Analysis
                                                                                                    2. RNA Sequencing (RNA-seq)
                                                                                                      1. Library Preparation
                                                                                                        1. Sequencing Platforms
                                                                                                          1. Bioinformatics Analysis
                                                                                                          2. Single-Cell RNA Sequencing
                                                                                                            1. Cell Capture Methods
                                                                                                              1. Data Processing
                                                                                                                1. Trajectory Analysis
                                                                                                              2. Functional Assays
                                                                                                                1. Colony-Forming Unit Assays
                                                                                                                  1. Hematopoietic CFU Assays
                                                                                                                    1. Mesenchymal CFU Assays
                                                                                                                    2. Differentiation Assays
                                                                                                                      1. Lineage-Specific Protocols
                                                                                                                        1. Functional Readouts
                                                                                                                        2. Migration and Invasion Assays
                                                                                                                        3. Teratoma Formation Assay for Pluripotency
                                                                                                                          1. Protocol and Interpretation
                                                                                                                            1. Histological Analysis
                                                                                                                              1. Alternative Pluripotency Tests
                                                                                                                              2. Epigenetic Analysis
                                                                                                                                1. Bisulfite Sequencing
                                                                                                                                  1. ChIP-Seq
                                                                                                                                    1. ATAC-Seq
                                                                                                                                    2. Live Cell Imaging
                                                                                                                                      1. Time-Lapse Microscopy
                                                                                                                                        1. Fluorescent Reporters
                                                                                                                                          1. Cell Tracking
                                                                                                                                        2. Genetic Manipulation
                                                                                                                                          1. Transfection and Transduction
                                                                                                                                            1. Methods
                                                                                                                                              1. Electroporation
                                                                                                                                                1. Lipofection
                                                                                                                                                  1. Viral Vectors
                                                                                                                                                  2. Efficiency and Safety
                                                                                                                                                    1. Transient vs. Stable Expression
                                                                                                                                                    2. Gene Editing Technologies
                                                                                                                                                      1. CRISPR-Cas9
                                                                                                                                                        1. Mechanism and Applications
                                                                                                                                                          1. Guide RNA Design
                                                                                                                                                            1. Off-Target Effects
                                                                                                                                                            2. TALENs
                                                                                                                                                              1. Mechanism and Applications
                                                                                                                                                                1. Design Principles
                                                                                                                                                                2. Zinc-Finger Nucleases
                                                                                                                                                                  1. Mechanism and Applications
                                                                                                                                                                    1. Protein Engineering
                                                                                                                                                                    2. Base Editing
                                                                                                                                                                      1. Cytosine Base Editors
                                                                                                                                                                        1. Adenine Base Editors
                                                                                                                                                                        2. Prime Editing
                                                                                                                                                                          1. Mechanism
                                                                                                                                                                        3. Lineage Tracing
                                                                                                                                                                          1. Genetic Labeling Techniques
                                                                                                                                                                            1. Cre-lox System
                                                                                                                                                                              1. Flp-FRT System
                                                                                                                                                                              2. Applications in Fate Mapping
                                                                                                                                                                                1. Clonal Analysis
                                                                                                                                                                                2. Reporter Systems
                                                                                                                                                                                  1. Fluorescent Proteins
                                                                                                                                                                                    1. Bioluminescence
                                                                                                                                                                                      1. Inducible Systems

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                                                                                                                                                                                  7. Applications and Therapeutic Potential