Application note Neuronal Networks with Molecular Devices
Karlijn Wilschut, Senior Scientist at Mimetas and Oksana Sirenko, Research Scientist at Molecular Devices wrote this application note on a quantitative, confocal high-content imaging method, enabling high-throughput phenotypic assessment of treatment effects upon the viability and morphology of 3D neuronal cultures in OrganoPlate®
Establishment of physiologically-relevant in vitro models is crucial to further understanding of the mechanisms of neurological diseases as well as targeted drug development. While iPSC-derived neurons show great promise for compound screening and disease modeling, use of three-dimensional (3D) cultures is emerging as a valid approach for neuronal cell based assay development. 3D cultures are recognized as more closely recapitulating aspects of the human tissues including the architecture, cell organization, as well as cell-cell and cell-matrix interactions.
The focus of this study is to develop a high-throughput 3D neurite outgrowth assay using iPSC-derived neurons developed in the OrganoPlate platform, with the goal of establishing 3D models for neurodegenerative diseases and neurotoxicology screens. The OrganoPlate is suitable for long- term culture of live cells, is amenable for screening purposes, and is compatible with standard laboratory equipment or automated systems, such the ImageXpress® Micro Confocal High-Content Imaging System.
Figure: iCell neurons plated into OrganoPlate capillary wells, in Matrigel, at 30,000 cells per chip. Cells were stained with the Calcein AM, MitoTracker Orange, and Hoechst. Images were taken using the ImageXpress Micro Confocal system