PT - JOURNAL ARTICLE AU - Katarina Bartalska AU - Verena Hübschmann AU - Medina Korkut-Demirbaş AU - Ryan John A. Cubero AU - Alessandro Venturino AU - Karl Rössler AU - Thomas Czech AU - Sandra Siegert TI - A systematic characterization of intrinsically formed microglia-like cells during retinal organoid differentiation AID - 10.1101/2021.12.30.474511 DP - 2022 Jan 01 TA - bioRxiv PG - 2021.12.30.474511 4099 - http://biorxiv.org/content/early/2022/01/01/2021.12.30.474511.short 4100 - http://biorxiv.org/content/early/2022/01/01/2021.12.30.474511.full AB - Brain organoids differentiated from human induced pluripotent stem cells provide a unique opportunity to investigate the development, organization and connectivity of neurons in a complex cellular environment. However, organoids usually lack microglia, brain-resident immune cells which are both present in the early human embryonic brain and participate in neuronal circuit development.Here, we find that microglia innately develop in unguided retinal organoid differentiation between week 3 and 4 in 2.5D culture and appear later in floating, non-pigmented, 3D-cystic compartments. We enriched for cystic structures using a low-dosed BMP4 application and performed mass spectrometry, thus defining the protein composition of microglia-containing compartments. We found that cystic compartments expressed both mesenchymal and epithelial markers with microglia enriched in the mesenchymal region. Interestingly, microglia-like cells started to express the border-associated macrophage marker CD163. The preferential localization of human microglia to a mesenchymal compartment provides insight into the behavior and migration of microglia. The model will ultimately allow detailed study of these enigmatic cells and how they enter and distribute within the human brain.Competing Interest StatementThe authors have declared no competing interest.