In this presentation, Gabriele Ciceri will introduce experimental strategies to generate neurons from human pluripotent stem cells via directed differentiation. Specifically, Ciceri will:

• Describe general principles of neuronal specification in vivo and their application to in vitro differentiation protocols.
• Introduce how to mimic development in a dish.

• Introduce paradigms for neural induction (PNS vs. CNS).
• Describe specification of regional identity through integration of multiple patterning signals and generation of specific neuron type.
• Explain how to control for lineage progression.
• Describe validation of directed differentiations.

Learning Objectives

After watching this presentation, participants at all career stages should be able to:

• Summarize the rationale of directed neuronal differentiation from human pluripotent stem cells.
• Describe how to apply the use of small molecules/morphogens to generate major neuron types.
• Introduce step-wise quality control criteria to assure the generation of the desired neuron type and eventually adjust conditions/troubleshoot the differentiations.

Supporting Resources

Download this guide to access resources that supplement Ciceri’s presentation.

Speaker

Gabriele Ciceri, PhD

Gabriele Ciceri is a research fellow at Memorial Sloan Kettering Cancer Center (MSKCC). He received his BS and MS in pharmaceutical biotechnology from the University of Milan, in Italy, and completed his MS thesis at San Raffaele Scientific Institute, also in Milan, in the laboratory of Ivan de Curtis. Prior to obtaining his PhD in neuroscience from the Miguel Hernández University, in Spain, he joined the laboratory of Oscar Marín at the Neuroscience Institute, in Alicante, Spain, to study the principles of lineage development and the generation of cell diversity in the mammalian cerebral cortex. He then joined the lab of Lorenz Studer in the Center for Stem Cell Biology at MSKCC as a postdoctoral fellow. Ciceri’s current research interests include the specification of CNS neuron identities from human pluripotent stem cells and defining the mechanisms controlling developmental timing of human neuronal maturation.

In this presentation, Nan Yang will:

• Introduce lineage conversion, a few principles on identifying transcription factors for different neuronal subtypes, and the application of the method.
• Describe detailed procedure of generating induced neuronal cells from human iPS cells and create neuron & astroglia co-cultures.
• Share troubleshooting tips for common problems.

Learning Objectives

After watching this presentation, participants at all career stages should be able to:

• Summarize the principles for direct lineage conversion.
• Identify the key techniques used to directly generate human neurons from iPS cells by transiently expressing transcription factors.
• Explain how to establish long-term culture for disease modeling.

Supporting Resources

Download this guide to access resources that supplement Yang’s presentation.

Speaker

Nan Yang, PhD

Nan Yang is an assistant professor of neuroscience at the Icahn School of Medicine at Mount Sinai. She is also a member of the Black Family Stem Cell Institute at Mount Sinai. Before she started her own lab in 2017, she was a postdoctoral trainee of Marius Wernig at Stanford University, where she pioneered the work in direct lineage reprogramming from somatic cells to neural cells and its application in disease modeling. She earned her PhD in genetics from Fudan University, in China. During her PhD training, she worked as a joint graduate student in Su Guo’s laboratory at the University of California, San Francisco, where she used zebrafish model organism to study the transcriptional signaling cascade that governs the differentiation of neural progenitor cells to dopaminergic neurons and interneurons in the basal forebrain region.

In this presentation, Zhiping Pang will introduce parameters for defining human neurons derived from iPSCs. Specifically, Pang will:

• Describe morphological features of human neurons (immature to mature characteristics).
• Introduce morphological and biochemical techniques to characterize human neurons.
• Provide information about functional characterization of human neurons, including electrophysiology (cover intrinsic neuronal membrane property; active membrane property; synapse transmission) and calcium imaging (network activity and contributions of synaptic transmission).

Learning Objective

After watching this presentation, participants at all career stages should be able to:

• Outline techniques and methods used to define human neurons derived from human iPS cells.

Supporting Resources

Download this guide to access resources that supplement Pang’s presentation.

Speaker

Zhiping Pang, PhD

Zhiping Pang is an associate professor of neuroscience and cell biology at Rutgers Robert Wood Johnson Medical School. Pang’s main research interests lie in understanding mechanisms of synaptic regulation from stem cells to the brain. Under the tutelage of Thomas Südhof, he earned his PhD in neuroscience from the University of Texas Southwestern Medical Center and completed postdoctoral training at Stanford University.