Fabrication and characterisation of random and aligned electrospun scaffolds to investigate hypothalamic stem/progenitor cell behaviour

Tanycytes are stem/progenitor cells that reside in the hypothalamus of the adult vertebrate brain. Tanycytes can be cultured as free-floating neurospheres in vitro but tend to spontaneously differentiate over time. Here we asked whether morphological cues provided by engineered polymer scaffolds can modify spontaneous differentiation. Tanycyte-derived neurospheres were cultured on electrospun scaffolds, prepared with either random or aligned fiber morphologies. Cells dispersed widely on the scaffolds, and - on aligned scaffolds - were highly organized, orientated parallel to the fibers. Immunocytochemical analysis showed that cells cultured on aligned scaffolds showed significantly greater expression of the neural stem/progenitor cell marker, NrCAM and reduced expression of differentiated cell markers in comparison to those cultured on random scaffolds. Together this shows that tanycytes respond to local engineered cues, and that a morphologically constrained environment can better maintain tanycytes as stem cells. The aligned scaffold culture system provides a powerful tool to better investigate this novel stem/progenitor cell population.