Developing neural organoids from IPSCs and observing neuronal and structural developmental through rt qPCR and immunofluorescence imaging
Neural organoids have proven to be an exemplary and novel method of modeling a number of congenital neurological anomalies, including rare childhood disorders associated with neurodevelopmental defects. Whereas traditional studies of human neurogenesis rely heavily on the use of animal models, organoids show promise in recapitulating and enacting the same human-derived physiological mechanisms. Here we investigated a way of generating dorsal and ventral organoids from IPSCs with a DOCK3 mutation to observe structural and developmental characteristics of neuronal differentiation through cryostaining and rt qPCR genomic expression. Immunofluorescent images generated through cyrostaining were able to detect the presence of Beta-Tubulin III, indicating neuronal development at day 15. Furthermore, rt qPCR genomic data revealed an upregulation of PAX6 in dorsal organoids, measuring early brain development. Organoid models show potential in modeling mutations of other neurological disorders to replicate pathophysiological mechanisms and cellular/molecular pathways manifested in clinical adaptations of these gene mutations.
