NPC-incorporated structured hydrogels to aid neural cell therapy

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Project Description

Structured hydrogels will specifically be compared with bulk hydrogel preparations with incorporated NPCs. The neural cells’ viability, differentiation/cell fate and distribution within the two types of gels will be evaluated.

Hypothesis to be tested: • Structured hydrogels offer a better support for human spinal cord derived NPCs compared to bulk hydrogel or 2D NPC culture as evaluated by the NPCs viability, cell fate potential and distribution patterns.

1. Cell viability of NPCs. (1) Light-sheet microscope overview (MIP) images of NPCs cultured in bulk and structured hydrogel (LIVE/DEAD viability assay will label all cells and dead cells). The 2D culture will be imaged with confocal. (2) Quantification and statistics of cell viability of different time points.

2. Neural differentiation of NPCs. Images of NPCs fluorescent staining in bulk and structured hydrogel (β-tubulin; GFAP). Analysis of cell differentiation (A) β-tubulin+ cell percentage; (B) GFAP+ cell percentage; (C) 3D reconstruction image/video to reflect the distribution of different cells (if possible, quantify the distance of the different cells to the surface of hydrogel and compare if the distances of the two types of cells to surface are different from each other); (D) Cell morphology (quantify neurite length/number of neurite process/number of neurite bearing cells, compare if this data from different hydrogels are different from each other).