Phenotyping of Human iPSC-derived Neurons
Patient-Driven Research
- 1st Edition - September 9, 2022
- Imprint: Academic Press
- Editor: Elizabeth D. Buttermore
- Language: English
- Paperback ISBN:9 7 8 - 0 - 1 2 - 8 2 2 2 7 7 - 5
- eBook ISBN:9 7 8 - 0 - 1 2 - 8 2 2 2 7 8 - 2
Phenotyping of Human iPSC-derived Neurons: Patient-Driven Research examines the steps in a preclinical pipeline that utilizes iPSC-derived neuronal technology to better understan… Read more
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Request a sales quotePhenotyping 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 presenting example projects that identify phenotypes and mechanisms relevant to autism spectrum disorder and epilepsy, this book allows readers to understand what considerations are important to assess at the start of project design. Sections address reproducibility issues and advances in technology at each stage of the pipeline and provide suggestions for improvement. From patient sample collection and proper controls to neuronal differentiation, phenotyping, screening, and considerations for moving to the clinic, these detailed descriptions of each stage of the pipeline will help everyone, regardless of stage in the pipeline.
In recent years, drug discovery in the neurosciences has struggled to identify novel therapeutics for patients with varying indications, including epilepsy, chronic pain, and psychosis. Current treatment options for such patients are decades old and offer little relief with many side effects. One explanation for this lull in novel therapeutics is a lack of novel target identification for neurological disorders (and target identification requires exemplar preclinical data). To improve on the preclinical work that often relies on rodent modeling, the field has begun utilizing patient-derived induced pluripotent stem cells (iPSCs) to differentiate neurons in vitro for preclinical characterization of neurological disease and target identification.
- Discusses techniques and new technology for iPSC culturing and neuronal differentiation to establish best practices in the lab
- Outlines considerations for phenotypic assay development
- Provides information about the successes, failures, and implications of phenotyping and screening with iPSC-derived neurons
- Describes how human iPSC-derived neurons are being used for preclinical discovery research as well as the development of therapeutics utilizing hiPSC-derived neurons
Academic and industry researchers in translational neuroscience and regenerative medicine
- Cover image
- Title page
- Table of Contents
- Copyright
- Dedication
- Contributors
- Section I. Best practices and considerations when designing a new project
- Chapter 1. iPSC culture: best practices from sample procurement to reprogramming and differentiation
- Facility setup
- Primary sample collection
- Reprogramming
- iPSC line characterization
- Best practices prior to differentiation
- Differentiation
- Chapter 2. Phenotypic assay development with iPSC-derived neurons: technical considerations from plating to analysis
- Introduction
- Establishing optimal conditions for phenotyping iPSC-derived neurons
- Assay development for screening
- Conclusion
- Chapter 3. Derivation of cortical interneurons from human pluripotent stem cells to model neurodevelopmental disorders
- Introduction
- A protocol for cortical interneuron derivation from human pluripotent stem cells (hPSCs)
- Enrichment and purification of cIN neural progenitor cells and neurons
- Cellular phenotyping of hPSC-derived cINs
- Alternate protocol for derivation of cIN NPCs from hPSCs
- Alternate protocol for differentiation of cIN NPCs into interneurons
- Chapter 4. Development of transcription factor-based strategies for neuronal differentiation from pluripotent stem cells
- Introduction
- Neuron differentiation driven by transcription factors
- Transcription factor-driven differentiation: considerations when designing a new protocol
- Summary and future directions
- Chapter 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
- Disease phenotyping of Purkinje cells
- Future perspectives for stem cell-derived Purkinje cells in translational medicine
- Chapter 6. Brain organoids: models of cell type diversity, connectivity, and disease phenotypes
- Introduction
- Cerebral organoids
- Other brain region specific organoids
- Neuronal activity and connectivity
- Non-neuronal cells
- Use of models in disease
- Reproducibility
- Conclusions and future directions
- Section II. The use of iPSC-derived neurons to study neurological disorders
- Chapter 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
- Conclusion
- Chapter 8. Use of cerebral organoids to model environmental and gene x environment interactions in the developing fetus and neurodegenerative disorders
- Introduction
- Maternal immune activation
- Cerebral organoids as a model system to study infectious diseases that cause neurodevelopmental disorders
- Cerebral organoids and cellular stress
- Cerebral organoids to model neurodegenerative disorders
- Conclusion
- Chapter 9. iPSC-derived models of autism: Tools for patient phenotyping and assay-based drug discovery
- Introduction
- Syndromic autisms
- iPSC studies to model ASDs in vitro
- 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
- Conclusions
- Chapter 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
- Neuronal networks as converging pathways?
- The way forward
- Chapter 11. Advantages and limitations of hiPSC-derived neurons for the study of neurodegeneration
- Introduction
- Biology of Alzheimer's disease
- Alzheimer's disease hiPSC models
- Tauopathies: Alzheimer’s disease related dementias
- The challenge of aging in hiPSC models of age-related disease
- Modeling Alzheimer's disease with cerebral organoids
- Using hiPSC models for drug discovery
- Conclusions
- Section III. New technology, industry perspective, and transitioning to the clinic
- Chapter 12. Developing clinically translatable screens using iPSC-derived neural cells
- Introduction
- Is an iPSC-derived platform right for the application?
- What factors should be considered in developing iPSC-based assays?
- What factors should be considered when running an iPSC-based screen?
- Summary
- Chapter 13. Gene editing hPSCs for modeling neurological disorders
- Introduction to limitations of iPSC-derived neuronal models that can be improved with gene editing
- Gene editing systems – past, present, and future
- Use of genetic modification to generate isogenic cell lines
- Safe harbor locus transgenic systems
- Endogenous locus transgenes
- Complex transgenic systems utilizing multiple gene editing events
- Future of genetic modification in hPSC-based neuronal research
- Chapter 14. Cell therapy and biomanufacturing using hiPSC-derived neurons
- Introduction
- hiPSC-derived neurons to model specific neurological disorders
- Brief history of bio-manufacturing
- Manufacturing hiPSCs and neuronal derivatives
- Clinical trials with hiPSC-derived neurons
- Perspective and challenges for clinical translation
- Chapter 15. Ethical considerations for the use of stem cell-derived therapies
- Overview of induced pluripotent stem cell (iPSC) therapies for neurological application
- Ethical and social issues
- Ethical translation of promising stem cell-based neurological therapeutics
- Conclusions
- Index
- Edition: 1
- Published: September 9, 2022
- No. of pages (Paperback): 372
- No. of pages (eBook): 372
- Imprint: Academic Press
- Language: English
- Paperback ISBN: 9780128222775
- eBook ISBN: 9780128222782
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Elizabeth D. Buttermore
Dr. Elizabeth Buttermore is currently the Director of Translational In Vitro Models in the Rosamund Stone Zander Translational Neuroscience Center at Boston Children’s Hospital (BCH) where she leads the Human Neuron Core and Neurological Repository Core. Elizabeth has 10 years of experience in phenotypic assay development using high content imaging and multielectrode array approaches. One of her goals at BCH is to help standardize the way the field obtains and interprets phenotypic data and to help researchers across academia and industry move their research forward. She completed her postdoctoral work in Clifford Woolf’s lab at BCH where she developed protocols for differentiating nociceptive neurons from human fibroblasts and iPSCs and used them to develop models for neuropathy and neuropathic pain. Prior to coming to BCH, Elizabeth completed her PhD in Neurobiology in 2012 at the University of North Carolina, in the lab of Manzoor Bhat, where she studied the organization and maintenance of molecular domains in myelinated axons.
Affiliations and expertise
Assistant Director for the Human Neuron Core at Boston Children’s Hospital (BCH), Phenotyping Services, USARead Phenotyping of Human iPSC-derived Neurons on ScienceDirect