Assessing inter-line reproducibility of a novel iPSC-derived 3D brain tissue model (3BTM) and comparison to 2D co-cultures

Stem-cell-based in vitro models offer promising potential to elucidate human brain cell functions and interactions under healthy conditions and in disease states. However, harnessing this potential is currently impaired by low reproducibility, maturity, and cell-type diversity of models. Therefore, we developed a novel 3D brain tissue model (3BTMs) containing neurons, astrocytes, and microglia with high reproducibility, maturity, and viability. To confirm reproducibility of culture generation, composition, and maturation across iPSC lines, and compare 3BTMs to 2D co-cultures of the same cells, we performed proteome analysis of 3BTMs from 3 independent wild-type cell lines over time and in comparison to 2D co-cultures, fetal and juvenile brain samples, and neural organoids from 2 independent lines.