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Defining the Role of Sonic Hedgehog in Neural Differentiation of Mouse Embryonic Stem Cells using Mass Cytometry

Presenters Name: 
Quoc-Tuan Nguyen
Co Presenters Name: 
Primary Research Mentor: 
Eli Zunder
Secondary Research Mentor: 
Kristen Fread
9:30 - 10:15
Time of Presentation: 
2019 - 9:30am to 10:15am
Newcomb Hall Ballroom
Presentation Type: 
Presentations Academic Category: 
Grant Program Recipient: 
Harrison Undergraduate Research Grant

Neurodegeneration of motor neurons can lead to neurological disorders, including ALS and progressive bulbar palsy. The role of sonic hedgehog (SHH) protein is well characterized in vivo, especially in the spinal cord to generate lower motor neurons, and may offer future therapeutic benefits. Previous studies have shown that added SHH during in vitro differentiation can generate Olig2 positive oligodendrocyte progenitor cells (OPCs) and HB9 positive motor neurons. We hypothesize that the increasing concentrations of SHH will generate an increased amount of OPCs and varying amounts of ventral neuron subtypes (such as floor plate neurons and upper motor neurons) depending on SHH concentration. Mass cytometry and FLOWMAP analysis allow for high dimension visualization of the generation of different neuronal cell types and can suggest lineage hierarchy relationships. Mouse embryonic stem cells (mESCs) were cultured in vitro and differentiated into neuronal lineage following a standard, 14-day N2B27 differentiation protocol. The concentration and time of addition of SHH to these neuronal differentiation cultures will be varied to investigate the role of SHH. The samples will be stained with metal conjugated antibodies and analyzed by single-cell mass cytometry to track neural surface markers and transcription factors. Preliminary results of time-course analysis of our N2B27 differentiation shows unique glial and neuronal populations in our differentiated culture. Comparing in vivo and in vitro studies may provide molecular insight into neurodevelopmental disorders and diseases.