The human brain is difficult to model in vitro due to the cellular diversity and thousands of
functional cellular interactions each neuron makes. In recent years, new advanced 3D cell
culture models known as cerebral organoids (COs) have emerged that more accurately reflect
the 3D structure and cellular diversity and interactions observed in the human brain.
However, the overwhelming majority of neurons within COs are seemingly arrested in
fetal-like states even after extended culture periods, which poses a major challenge to their
utility for modelling disorders and diseases of the postnatal human brain. To overcome this
issue of limited maturation, we have predicted transcription factors that potentially regulate
neuronal maturation using our lab's recently published brain development cell atlas1, which
profiled the transcriptional (snRNA-seq) and epigenetic states (snATAC-seq) of individual
nuclei in the human prefrontal cortex from mid gestation to adulthood. A single nucleus TF
screen will be performed in COs to identify TF combinations that enhance maturation the
most, using the brain development cell atlas snRNA-seq dataset as a maturation state
reference. Additionally, the effectiveness of several other approaches to enhance maturation
in COs such as, fusing brain region specific organoids, using optogenetics to stimulate
neuronal activity, and small molecule treatments will be benchmarked using snRNA-seq to
directly compare to the developing human prefrontal cortex. By enhancing maturation, COs
will more closely resemble the postnatal human brain, which will be highly advantageous for
disease modelling or drug screening approaches for diseases that onset later in life.
Furthermore, approaches that enhance maturation in COs might be applicable to the many
other human tissue cell culture models (e.g. cardiomyocytes) that also face this issue of
limited maturation.