Neuron Culture

Figure 1. CytoGrow™ growth factors support 3D encapsulation culture of NSC cells as single cell suspension in VitroGel® NEURON using RocketCell™ NSC Xeno-Free Growth Medium. ​

VitroGel® NEURON Hydrogel and RocketCell™ NSC Xeno-Free Media Support 3D NSC Cultures and Neurite Outgrowth. The growth medium was supplemented with CytoGrow EGF and FGF2. 3D NSC cultures were established by preparing a single-cell NSC suspension with RocketCell™ NSC Xeno-Free Medium and mixing with VitroGel NEURON for 6 days (A). Immunofluorescence staining was performed to evaluate the presence of beta-III-tubulin, a neuron maturation marker. The images were obtained using the Keyence BZ-X microscope at 10x and 20x magnifications (B).

Figure 2. CytoGrow™ growth factors support NSC growth and maintenance on VitroGel® NEURON coating surface of VitroPrime™ Spread-Attach using RocketCell™ NSC Xeno-Free Growth Medium. ​

24-well plates were coated with VitroGel® NEURON hydrogel (1:200 dilution) for 30 minutes at room temperature. iPSC-derived NSCs (4 x 104 cells/well) were added to each well, resulting in a final volume of 0.5 mL per well. The medium was changed every 2 days, and the cultures were monitored using an inverted microscope at a 10X magnification. ​

Figure 3. Confocal imaging of IPSC spheroids growing in VitroGel®-STEM.

IPSC cultures were plated in VitroGel®-STEM and grown for 5 days. The slides were imaged using a Leica Mica confocal microscope. An IPSC spheroid was imaged in 3D using the Z-stacking function, and images for Hoechst 33342 (DNA), Tra-1-60 (Podocalyxin, cell surface), Lin28A (cytoplasmic RNA binding protein), and Oct4 (nuclear transcription factor) were projected using the maximum intensity algorithm. ​