In this study, we aim to develop new platforms for neurosphere assay and single cell manipulation for neural stem cell research. Neurosphere assay is a common method for identification of neural stem/progenitor cells, but obtaining single cells from dissociated neurospheres is difficult using non-enzymatic methods. We developed a microfluidic-chip-based approach that utilizes flow and microstructures to dissociate neurospheres. Results show that this microfluidic-chip-based neurosphere dissociation method can generate high yields of single cells from dissociated neurospheres of mouse KT98 and DC115 cell models for 90% and 95%, respectively. The microfluidic-chip-dissociated cells had high viabilities (80 - 85%) and the ability to re-grow into neurospheres, demonstrating the applicability of this device to neurosphere-assay applications. In addition, the dissociated cells retained their normal differentiation potentials, as shown by their capabilities to differentiate into three neural lineages (neurons, astroglia, and oligodendrocytes) when cultured in differentiation culture conditions.
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