Fluorescent brain organoids for the study of function and disease
November 2020
Ruhr University Bochum, Bochum, Germany
Cerebral organoids are a promising model to study human brain function and disease, but the high variability between organoids presents a challenge. A method using labelled mixed organoids generated from two different human iPSC (hiPSC) lines is presented. This allows the identification of cells of different origins within a single organoid and overcomes the limitations of inhomogeneity between organoids. Using a CRISPR/Cas9 gene-editing approach, different fluorescent proteins were fused to β-actin or lamin B1 in hiPSCs and subsequently used as markers to identify each cell line. Mixtures of differentially edited cells were seeded to induce embryoid body formation and cerebral organoid differentiation. Subsequently, 3D tissue development could be followed by confocal fluorescence microscopy and immunofluorescence staining in fixed samples. Overall, the results support the possibility of using differentially labelled reporter cell lines with different characteristics (e.g., mutation-containing and "healthy cells") to generate more accurate human models that allow direct comparison under identical experimental conditions.
Gene edited fluorescent cerebral organoids to study human brain function and disease
Thorsten Müller
Added on: 05-12-2021
[1] https://www.biorxiv.org/content/10.1101/2020.11.24.395533v1
