3D Analysis for 3D Cell Models: A Method for Assessing Mitotic Spindle Orientation in Cerebral Organoids
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2022-10-04
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en
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Oriented division of the mitotic spindle is a highly regulated and evolutionarily conserved process that is critical to proper tissue development. Disruption of the mitotic spindle has been implicated as a mechanism behind neurodevelopmental diseases such as microcephaly. Especially in early radial glia progenitor cells, divisions with a spindle orientation that is parallel to the surface of the cerebral ventricles has been found to be predictive of proliferative symmetric division, while perpendicular spindle orientations are predictive of differentiative asymmetric division, making the analysis of spindle orientation an informative assay. However, research which aims to use the parameter of mitotic spindle orientation to characterize the effects of a genetic anomaly on the development of an in vitro or in vivo model often utilizes a 2D method of analysis that has been found to have significant systematic biases. Here we introduce an adapted method for assessing mitotic spindle orientation in 3D which works for any 3D cell or animal model. This method is more accurate and quicker to perform than the 2D method mentioned earlier and is cheaper and easier for many labs to implement than alternative methods such as live-cell imaging. We also demonstrate the use of our method to assess mitotic spindle orientation defects in Koolen de Vries Syndrome cerebral organoids and find that haploinsufficiency of KANSL1 results in a decrease in planar divisions at early developmental timepoints.
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Faculteit der Sociale Wetenschappen