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Old URL: https://wesscholar.wesleyan.edu/etd_hon_theses/1889
Drupal Node ID: 23848
author email: cdundes@wesleyan.edu
old filename: 1169_242768_1889"},{"label":"Degree","value":["Bachelor of Arts with Departmental Honors"]},{"label":"Description","value":"

Human embryonic stem cell-derived neural progenitors (hESNPs) can be used for cell replacement therapy in neurodegenerative diseases such as temporal lobe epilepsy (TLE). We use hESNPs to derive inhibitory interneurons lost in the hippocampal region of the brain in patients with TLE to test the efficacy of establishing a transplant therapy to reduce seizures in an animal model. Mature inhibitory interneurons produce the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). These interneurons predominantly arise in vivo from a transient embryonic structure, the medial ganglionic eminence (MGE), which is characterized by the expression of the transcription factor NKX2.1. Prominent subtypes of inhibitory interneurons derived from the MGE express either parvalbumin (PV) or somatostatin (SST), and these subtypes are specifically affected in TLE. However, we have had limited success in generating hESNP-derived PV-expressing subtypes. In this thesis, we seek to optimize culture conditions for the generation of GABA-positive interneuron subtypes through adjustments to the traditional in vitro astrocyte co-culture system and experimentation with an ex vivo organotypic hippocampal slice culture model. We find that hESNPs differentiated in long-term co-culture express interneuron subtype markers and indicate functional phenotypes through their ability to fire trains of action potentials. Additionally, our data indicate that hESNPs grafted on ex vivo hippocampal slices generate mature GABAergic interneuron subtypes at higher levels than found in astrocyte co-culture. This research provides evidence for the robust differentiation potential of hESNPs into functional subtypes of GABAergic inhibitory interneurons through two culture systems modeling in vivo development.

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