Phenotyping of human IPSC-derived neurons : patient-driven research /
"Phenotyping of Human iPSC-derived Neurons: Patient-Driven Research examines the steps in a preclinical pipeline that utilizes iPSC-derived neuronal technology to better understand neurological disorders and identify novel therapeutics, also providing considerations and best practices. By prese...
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| Format: | eBook |
| Language: | English |
| Published: |
London ; San Diego, CA :
Academic Press, an imprint of Elsevier,
[2023]
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| Subjects: | |
| Online Access: | Connect to the full text of this electronic book |
Table of Contents:
- Front Cover
- PHENOTYPING OF HUMAN IPSC-DERIVED NEURONS
- PHENOTYPING OF HUMAN IPSC-DERIVED NEURONS: PATIENT-DRIVEN RESEARCH
- Copyright
- Dedication
- Contents
- Contributors
- I
- Best practices and considerations when designing a new project
- 1
- iPSC culture: best practices from sample procurement to reprogramming and differentiation
- Facility setup
- Tissue culture room design
- Tissue culture equipment
- Primary sample collection
- Somatic cells
- Quality control of somatic cells
- Reprogramming
- Pros and cons of each method
- Episomal vector transfection
- Sendai virus transduction
- mRNA reprogramming method
- iPSC line characterization
- Sterility
- Pluripotency
- Transgene elimination
- Identity
- Genetic stability
- Best practices prior to differentiation
- Cell banking
- Culturing conditions
- Differentiation
- Experimental design
- Cell line selection
- Differentiation protocol selection
- Best practices during differentiation
- References
- 2
- Phenotypic assay development with iPSC-derived neurons: technical considerations from plating to analysis
- Introduction
- Establishing optimal conditions for phenotyping iPSC-derived neurons
- Differentiation protocol considerations
- Coating substrates
- High content imaging (HCI)
- Functional analysis
- Multi-electrode array (MEA) recording
- Calcium imaging
- Patch clamping
- Live imaging
- Fluorescent microplate assays
- Assay development for screening
- Conclusion
- References
- 3
- Derivation of cortical interneurons from human pluripotent stem cells to model neurodevelopmental disorders
- Introduction
- Development of the human cortex
- Modeling human cortical interneuron development in vitro
- The development of protocols for cortical interneurons from human pluripotent stem cells (hPSCs) to model neurodevelopmenta.
- A protocol for cortical interneuron derivation from human pluripotent stem cells (hPSCs)
- Equipment and supplies
- Reagents
- Preparation of reagents
- Accutase cell detachment solution
- B-27 supplement (50×) minus vitamin A
- Preparing matrigel
- Coating tissue culture plates with Matrigel
- Coating tissue culture plates with Matrigel-Laminin
- Small molecule preparation
- Media composition
- Protocol
- Specification of cortical interneuron progenitors from hPSCs
- Maintenance and expansion of cIN NPCs
- Cryopreservation of cIN neural progenitor cells
- Revival and maintenance of cryopreserved cIN neural progenitor cells
- Interneuron differentiation and maturation from cIN neural progenitor cells
- Enrichment and purification of cIN neural progenitor cells and neurons
- Enrichment for post-mitotic cINs with neural rosette selection reagent
- Purification of post-mitotic cINs with NCAM bead selection
- Critical steps and troubleshooting
- Cellular phenotyping of hPSC-derived cINs
- Using immunocytochemistry to benchmark hPSC-derived cINs and to assess NDD-related alterations of neurodevelopment
- Morphometric analysis of neurite extension and length
- Neuronal migration assay
- Measurement of synaptic puncta
- Alternate protocol for derivation of cIN NPCs from hPSCs
- Alternate protocol for differentiation of cIN NPCs into interneurons
- Acknowledgments
- References
- 4
- Development of transcription factor-based strategies for neuronal differentiation from pluripotent stem cells
- Introduction
- Neuron differentiation driven by transcription factors
- Dopaminergic (DA) neurons
- Glutamatergic neurons
- GABAergic neurons
- Cholinergic motor neurons
- Retinal ganglion cells
- Glia: astrocytes, oligodendrocytes, and microglia.
- Transcription factor-driven differentiation: considerations when designing a new protocol
- Design a cocktail of transcription factors
- Transcription factor delivery
- Genome integrating vectors
- Non-genome integrating viral vectors
- Synthetic mRNA
- Summary and future directions
- Acknowledgement
- References
- 5
- Differentiation of Purkinje cells from pluripotent stem cells for disease phenotyping in vitro
- Development of the cerebellum
- Differentiation of pluripotent stem cells into Purkinje cells
- Cerebellar organoids derived from iPSCs and ESCs in 3D cultures
- Human iPSC- and ESC-derived Purkinje cell differentiation in 2D co-cultures with mouse cerebellar cells
- Functional characterization of human pluripotent stem cell-derived Purkinje cells in vitro and in vivo
- Challenges in the differentiation of human Purkinje cells in 2D- and 3D-cell cultures
- Disease phenotyping of Purkinje cells
- Purkinje cells in cerebellar ataxia
- Mouse Purkinje cell models of cerebellar ataxia
- Human iPSC-derived NPCs and Purkinje cells in cerebellar ataxia
- Purkinje cells in Tuberous Sclerosis Complex (TSC)
- Mouse Purkinje cell models of TSC
- TSC patient iPSC-derived Purkinje cells
- Future perspectives for stem cell-derived Purkinje cells in translational medicine
- Cell transplantation for treatment of cerebellar degeneration
- Drug screening with pluripotent stem cell-derived Purkinje cells
- Acknowledgments
- References
- 6
- Brain organoids: models of cell type diversity, connectivity, and disease phenotypes
- Introduction
- Cerebral organoids
- Human corticogenesis overview
- Organoid differentiation overview
- Fidelity of hCO cell types and organization
- Other brain region specific organoids
- Neuronal activity and connectivity
- Synaptic activity
- Connectivity of neuronal organoids
- Non-neuronal cells.
- Astrocytes
- Oligodendrocytes
- Microglia
- Vascularization/nutrient distribution
- Summary of non-neuronal cells
- Use of models in disease
- Microcephaly modeling with hCOs
- ASD modeling with hCOs
- Molecularly defined ASD
- Idiopathic ASD
- Limitations of hCO modeling for CNS disorders
- Reproducibility
- Sources of variability in organoid model systems
- Addressing reproducibility
- Conclusions and future directions
- References
- II
- The use of iPSC-derived neurons to study neurological disorders
- 7
- Human models as new tools for drug development and precision medicine
- Introduction
- Drug development pipeline
- Human models as a screening tool for personalized precision medicine
- Monolayer models
- Organoids
- Organ-on-chip platforms
- Conclusion
- References
- 8
- Use of cerebral organoids to model environmental and gene x environment interactions in the developing fetus an ...
- Introduction
- Maternal immune activation
- Cerebral organoids as a model system to study MIA and neuroinflammation
- Cerebral organoids as a model system to study infectious diseases that cause neurodevelopmental disorders
- Zika virus
- SARS-CoV-2
- Human immunodeficiency virus (HIV)
- Toxoplasmosis
- Cytomegalovirus (CMV)
- Herpes simplex virus (HSV)
- Cerebral organoids and cellular stress
- Heat shock
- Fetal alcohol syndrome
- Cerebral organoids to model neurodegenerative disorders
- Alzheimer's disease (AD)
- Cerebral organoids in familial AD
- Modeling sporadic AD
- Cerebral organoids for drug development in AD
- Modeling Parkinson Disease using organoid cultures
- Conclusion
- References
- 9
- iPSC-derived models of autism: Tools for patient phenotyping and assay-based drug discovery
- Introduction
- Syndromic autisms
- Fragile X syndrome
- Rett syndrome
- FOXG1 deletion syndrome
- Tuberous sclerosis.
- Pheland McDermid syndrome
- Prader-Willi and Angelman syndromes
- Timothy syndrome
- iPSC studies to model ASDs in vitro
- iPSC studies focused on syndromic and sporadic autisms
- iPSC studies focusing on sporadic non-syndromic autism
- Data collected by studies focused on iPSCs from idiopathic autism
- Gene expression profiling
- Concordances in gene expression profiles obtained from studies on iPSC-derived cells and post-mortem brain tissue from idio ...
- Morphological and electrophysiological properties in iPSC-derived neurons from patients with idiopathic autism
- Similar phenotypes between iPSC-derived neurons from patients with sporadic or syndromic autisms and idiopathic autism
- 3D models of ASDs-a focus on organoids, spheroids, and assembloids
- The use of iPSCs to develop assays and novel therapies that can be translated to the clinic for ASD
- Limitations for using iPSC-derived neurons in drug screening platforms
- Quality control testing
- Automation challenges
- Cost
- Small "n"
- Epigenetic memory
- Well-to-well variability
- Variability within cell lines
- Variability across differentiation batches
- Disease modeling
- Screening of simple phenotypes
- The use of iPSC-derived neurons for personalized medicine
- Conclusions
- References
- 10
- Probing the electrophysiological properties of patient-derived neurons across neurodevelopmental disorders
- Induced pluripotent stem cells and modeling brain disorders
- Progressing from gene discovery to functional gene groupings to pathophysiology
- Neuronal networks represent a logical level for the manifestation of NDDs
- Micro-electrode arrays as a scalable high-throughput functional assay
- Phenotyping NDD patient-derived neurons using MEA recordings
- Fragile X and Rett syndrome
- Kleefstra syndrome
- Neuronal networks as converging pathways?.