Recently, 3D cell culture neural disease models have become attractive alternatives to animal models and traditional 2D cell cultures. Co-cultures of neurons, astrocytes and oligodendrocytes have been shown to recapitulate many in vivo CNS phenotypes including neural synaptic complexity, gene expression patterns and neuroinflammatory responses. However, these models often rely on undefined animal derived hydrogels which are not amendable to high throughput screening applications. The TrueGel3D® HTS Hydrogel Plate is a ready-to-use solution to easily establish 3D cell cultures using fully synthetic hydrogels in a simple and automation-compatible manner. Here we demonstrate how ReNcell® VM human neural stem cells can be differentiated into β-tubulin expressing neurons using TrueGel3D® HTS hydrogel plates following an optimized step-by-step culture protocol.
ReNcell® VM Human Neural Progenitor Cell Line (SCC008) can be expanded on laminin (CC095-M) coated (20 µg/mL) tissue culture plastic first and then seeded in TrueGel3D® HTS Hydrogel Plates (TRUE-HTS1, TRUE-HTS10). Alternatively, cells can be thawed and directly seeded in TrueGel3D® HTS Hydrogel Plate. Ensure cells are at high viability before seeding into TrueGel3D® HTS Hydrogel Plates. The cells should be handled and stored according to the manufacturer’s instructions. All steps should be performed inside a laminar flow hood to ensure sterility.
Figure 1. Neural Differentiation of Human NSCs using TrueGel3D® HTS Hydrogel Plates.
Fluorescent staining of day 12 and 23 neural differentiated ReNcell® VM NSC cultured in 3D using TrueGel3D® HTS hydrogel plates acquired with an Olympus SpinSR10 spinning disk confocal microscope. Neurons express β-Tubulin III (Red) and counterstained with Hoechst 3358 (Blue).
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