Module 3: Generating Glial and Mixed Cultures
Learn how to produce and characterize three major glial cell types in the brain - oligodendrocytes, astrocytes, and microglia – and how to assemble co-cultures.
 Mathew Blurton-Jones, PhD University of California, Irvine |
 Hiroko Nobuta, PhD Albert Einstein College of Medicine |
 Jason Tchieu, PhD Memorial Sloan Kettering Cancer Center |
After reviewing Module 3, participants at all career stages should be able to:
- Outline basic methods that can be used to differentiate iPS cells into microglia and to validate the resulting phenotype.
- Identify assays that can be used to study the function of microglia in vitro and the potential use of chimeric transplantation models to study human microglia function in vivo.
- Describe growth factor-mediated human oligodendrocyte generation from iPS cells.
- Explain how to troubleshoot common experimental challenges.
- Explain the logic of directed differentiation of astrocytes from human pluripotent stem cells (hPSC’s).
- Describe some uses of astrocytes for modeling neuron/glia interactions in mixed cultures.
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Module 3A: Generation and Use of iPS-Derived MicrogliaIn this presentation, Mathew Blurton-Jones will:
- Introduce methods to differentiate pluripotent stem cells into microglia.
- Describe approaches that can be used to validate the resulting cells.
- Describe in vitro assays that can be used to examine microglial function.
- Identify potential caveats with in vitro microglial culture.
- Explain development and use of new chimeric mouse models to study human microglial function in vivo.
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Module 3B: Generation of Oligodendrocytes from iPS CellsIn this presentation, Hiroko Nobuta will introduce various methods for generating oligodendrocytes from iPS cells. Specifically, Nobuta will:
- Describe various methods of generating mouse and human oligodendrocytes and their pros and cons.
- Introduce the detailed procedure of the growth factor-mediated human oligodendrocyte generation from iPS cells.
- Provide troubleshooting tips for common problems in the growth factor-mediated human oligodendrocyte generation and functional assays.
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Module 3C: Deriving Human Astrocytes from Pluripotent Stem CellsIn this presentation, Jason Tchieu will:
- Explain why human astrocytes are important.
- Provide general methods to differentiate astrocytes from PSCs, including transcription factor mediated systems.
- Describe how to stage and troubleshoot the differentiation(s).
- Introduce functional and molecular characterization.
- Introduce co-culture analysis.
Module 3A: Generation and Use of iPS-Derived Microglia
In this presentation, Mathew Blurton-Jones will:
- Introduce methods to differentiate pluripotent stem cells into microglia.
- Describe approaches that can be used to validate the resulting cells.
- Describe in vitro assays that can be used to examine microglial function.
- Identify potential caveats with in vitro microglial culture.
- Explain development and use of new chimeric mouse models to study human microglial function in vivo.
Learning Objectives
After watching this presentation, participants at all career stages should be able to:
- Outline basic methods that can be used to differentiate iPS cells into microglia and to validate the resulting phenotype.
- Identify a variety of assays that can be used to study the function of microglia in vitro and the potential use of chimeric transplantation models to study human microglia function in vivo.
Supporting Resources
Download this guide to access resources that supplement Blurton-Jones’ presentation.
Speaker
Mathew Blurton-Jones, PhD
Mathew Blurton-Jones is an associate professor in the department of neurobiology and behavior at the University of California, Irvine, and director of UCI’s Alzheimer’s disease iPSC and CRISPR cores. His research combines patient-derived pluripotent stem cells, CRISPR gene-editing, and chimeric mouse models to examine the impact of Alzheimer’s disease risk genes on human microglial function. Blurton-Jones earned his PhD in neuroscience at the University of California, San Diego and completed postdoctoral training at UC Irvine before starting his own lab in 2011.
Module 3B: Generation of Oligodendrocytes from iPS Cells
In this presentation, Hiroko Nobuta will introduce various methods for generating oligodendrocytes from iPS cells. Specifically, Nobuta will:
- Describe various methods of generating mouse and human oligodendrocytes and their pros and cons.
- Introduce the detailed procedure of the growth factor-mediated human oligodendrocyte generation from iPS cells.
- Provide troubleshooting tips for common problems in the growth factor-mediated human oligodendrocyte generation and functional assays.
Learning Objectives
After watching this presentation, participants at all career stages should be able to:
- Describe growth factor-mediated human oligodendrocyte generation from iPS cells.
- Explain how to troubleshoot common experimental challenges.
Supporting Resources
Download this guide to access resources that supplement Nobuta’s presentation.
Speaker
Hiroko Nobuta, PhD
Hiroko Nobuta is a postdoctoral fellow in the neuroscience department at the Albert Einstein College of Medicine. She completed training in oligodendrocyte biology with her PhD mentor James Waschek, at the University of California, Los Angeles, as well as her postdoctoral mentors David Rowitch, at the University of California, San Francisco, and Marius Wernig, at Stanford University. She is now exploring gene engineering methods to manipulate glia cells to treat neurodegenerative disorders.
Module 3C: Deriving Human Astrocytes from Pluripotent Stem Cells
In this presentation, Jason Tchieu will:
- Explain why human astrocytes are important.
- Provide general methods to differentiate astrocytes from PSCs, including transcription factor mediated systems.
- Describe how to stage and troubleshoot the differentiation(s).
- Introduce functional and molecular characterization.
- Introduce co-culture analysis.
Learning Objectives
After watching this presentation, participants at all career stages should be able to:
- Explain the logic in the directed differentiation of astrocytes from hPSCs, including steps where transgene expression can bypass particular stages of neural development.
- Outline some uses of astrocytes for modeling neuron/glia interactions in mixed cultures.
Supporting Resources
Download this guide to access resources that supplement Tchieu’s presentation.
Speaker
Jason Tchieu, PhD
Jason Tchieu is a postdoctoral research fellow at Memorial Sloan Kettering Cancer Center. He previously worked as a bioinformatics programmer at the San Diego Supercomputer Center. Tchieu’s research focuses on understanding how the brain develops by elucidating the role of glia in neurological disorders. He received his undergraduate degree in biochemistry from the University of California, San Diego and his PhD in molecular biology from the University of California, Los Angeles.
