UC San Diego

Cellular models of brain disease involve genetic modulation, geometric patterning, neurophysiologic monitoring and analyses of both primary and immortalized cell lines. Additionally, recent neurological disease models often necessitate in vitro directed differentiation and maturation of human stem cell lines. To advance human stem cell based neural disease models within this evolving field, adaptive approaches of progressive complexity are essential. First, I invented an adaptable 3D laminar scaffold fabrication method to facilitate complex in vitro cell and tissue cultures. Next, we built on the Muotri Laboratory’s pioneering iPSC approach, incorporating emergent multi-well microelectrode array technology to establish the first functional in vitro human neural model of Cockayne Syndrome (CS). We revealed previously unreported neural network abnormalities and identified neuronal transcription dysregulation of the GH/IGF-1 signaling pathway. We also enhanced our previously established Rett Syndrome (RTT) model using advanced stem cell methods, including genomic editing technology to create isogenic matched MECP2 cell lines and establish neural co-cultures. With this system I revealed adverse astroglial non-autonomous effects on neuronal networking in monolayer co-cultures. Finally, we combined my novel multilayer fabrication method with this isogenic MECP2 approach to pattern neural co-cultures in 3D hyaluronic acid based laminar tissue scaffolds, building further complexity into our original model. We report accelerated in vitro neural development, NPC chemotaxis defects and additional astroglial non-autonomous effects in this new 3D biomimetic RTT neural model. The methods described herein have allowed us to establish multidimensional individualized human donor-relevant models of a neurodegenerative disorder (CS) and a neurodevelopmental disorder (RTT) in screenable formats.