Year 2

The use of stem cell technology to study neurodegenerative diseases has been a burgeoning area of research in recent years. Recent work by us and others have demonstrated that stem cell derived cortical neurons (CN) from Alzheimer’s disease patients demonstrate key disease differentials when compared to CN’s derived from healthy controls. However, most of this work has been limited to the use of simple cultures of iPSC-derived cortical or motor neurons, or in some cases, co-culture of iPSC-derived neurons with primary human astrocytes. We have demonstrated that co-culturing of iPSC-derived neurons with primary fetal astrocytes facilitates the maturation of neurons (as assessed by electrophysiology) beyond what is observed in neuronal cultures alone. We have developed an astrocyte differentiation protocol from iPSC that allows for the generation of cells with gene expression, glutamate uptake capacity and kinetics, and neurotropic factor secretion consistent with primary fetal astrocytes. Importantly, these iPSC astrocytes can be directed to an anterior versus posterior (brain versus spinal cord) identity. The goal of this study is to compare the ability of iPSC-derived astrocytes to primary fetal astrocytes in their ability to facilitate the maturation of iPSC-derived neurons, in order to identify the appropriate co-culture conditions where iPSC-derived neurons reach functional maturity as assessed by neuronal markers and electrophysiological activity. Achievement of this goal would allow for further assessment of the contribution of neurons and astrocytes to specific disease processes in neurodegeneration and neuroinflammation through the ability to combine in co-culture either disease versus healthy iPSC-derived astrocytes and/or neurons to further elucidate the underlying mechanisms contributing to these disease phenotypes. These studies should facilitate the development of co-culture systems which drive neuronal development to a more mature phenotype, and improve the availability of stem cell-based model systems to better replicate in vivo physiological and pathological processes.