AUTHOR=Eigenhuis Kristel N. , Somsen Hedda B. , van der Kroeg Mark , Smeenk Hilde , Korporaal Anne L. , Kushner Steven A. , de Vrij Femke M. S. , van den Berg Debbie L. C. TITLE=A simplified protocol for the generation of cortical brain organoids JOURNAL=Frontiers in Cellular Neuroscience VOLUME=17 YEAR=2023 URL=https://www.frontiersin.org/journals/cellular-neuroscience/articles/10.3389/fncel.2023.1114420 DOI=10.3389/fncel.2023.1114420 ISSN=1662-5102 ABSTRACT=

Human brain organoid technology has the potential to generate unprecedented insight into normal and aberrant brain development. It opens up a developmental time window in which the effects of gene or environmental perturbations can be experimentally tested. However, detection sensitivity and correct interpretation of phenotypes are hampered by notable batch-to-batch variability and low reproducibility of cell and regional identities. Here, we describe a detailed, simplified protocol for the robust and reproducible generation of brain organoids with cortical identity from feeder-independent induced pluripotent stem cells (iPSCs). This self-patterning approach minimizes media supplements and handling steps, resulting in cortical brain organoids that can be maintained over prolonged periods and that contain radial glial and intermediate progenitors, deep and upper layer neurons, and astrocytes.