A simplified protocol for the generation of cortical brain organoids

Kristel N Eigenhuis; Hedda B Somsen; Mark van der Kroeg; Hilde Smeenk; Anne L Korporaal; Steven A Kushner; Femke M S de Vrij; Debbie L C van den Berg

doi:10.3389/fncel.2023.1114420

A simplified protocol for the generation of cortical brain organoids

Front Cell Neurosci. 2023 Apr 4:17:1114420. doi: 10.3389/fncel.2023.1114420. eCollection 2023.

Authors

Kristel N Eigenhuis¹, Hedda B Somsen¹, Mark van der Kroeg², Hilde Smeenk², Anne L Korporaal¹, Steven A Kushner^{2

3}, Femke M S de Vrij², Debbie L C van den Berg¹

Affiliations

¹ Department of Cell Biology, Erasmus MC, Rotterdam, Netherlands.
² Department of Psychiatry, Erasmus MC, Rotterdam, Netherlands.
³ Department of Psychiatry, Columbia University Irving Medical Center, New York, NY, United States.

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.

Keywords: cortical brain organoids; dorsal identity; feeder-independent; human brain development; induced pluripotent stem cells; self-patterning.

Grants and funding

DB is supported by a Horizon 2020 Marie Curie Individual Fellowship (#799214) and an Erasmus University Rotterdam Fellowship. FV is supported by the Erasmus MC Human Disease Model Award. FV and SK are supported by the Netherlands Organ-on-Chip Initiative, an NWO Gravitation project (024.003.001) funded by the Ministry of Education, Culture, and Science of the Government of the Netherlands.