(1141-B) Developing a High-throughput Screening Assay for Myelination and Demyelination Promoting Compounds using Human iPSC-derived Schwann cell progenitor cells, Motor Neurons and Sensory Neurons from the Same Donor
Monday, February 5, 2024
2:00 PM – 3:00 PM EST
Abstract: Schwann cells are the primary glia within the peripheral nervous system where they associate with and myelinate both sensory neurons and motor neurons. These three cell types are implicated in various human diseases – including diabetic peripheral neuropathy, Charcot-Marie-Tooth syndrome, and pain – yet these diseases have been difficult to study in a human-relevant context due to lack of well characterized high-throughput translational in-vitro models. Here we demonstrate the rapid and efficient production of Schwann cell precursors (SCPs) from human induced pluripotent stem cells using directed differentiation under defined media conditions in 10 days. These SCPs were characterized via immunocytochemistry and qPCR for the markers OCT6, SOX10, S100B, and KROX20. To develop complex models of peripheral neurobiology, SCPs were co-cultured with sensory neurons and motor neurons derived from the same human induced pluripotent stem cell line using 7 day directed differentiation protocols. SCP-axon alignment was showcased after only one week in vitro, and myelination markers myelin basic protein and myelin protein zero were analyzed at multiple timepoints. Transmission electron microscopy revealed the ultrastructural details of Schwann cells engaging in the myelination of neuronal axons. Co-cultures were screened in dose response to tool compounds for myelination promotion (clemastine, tamoxifen, and T3) and de-myelination (cuprizone and lysophosphatidylcholine). Immunocytochemistry quantification of MBP and MPZ as well as morphological readouts were used to access compound effects. Functional characterization of the co-cultures using multi-electrode arrays were also explored. In summary, the ready availability of these hPSC-derived SCPs and sensory/motor neurons will rapidly advance the development of more high-throughput physiological models for human peripheral nervous system diseases.