Influence of Initial Seeding Density on Gene Expression during Neuronal Priming

Abstract

Derivation of neurons from pluripotent stem cells has moved from a niche expertise to a widely used technology, prompting a surge to create novel and timely treatments for neurologic disorders such as Parkinson's disease. However, current methods for derivation of neurons, though widely available, remain inefficient, resulting in sub-optimal yields and purities of potentially therapeutic cells. Optimising cell processing methods to improve yield and purity is necessary and in this work, we assessed the impact of Initial Cell Seeding Density (ICSD) on differentiation of pluripotent cells (human embryonic and human induced pluripotent stem cells) towards a neuronal specification using a recognised laboratory protocol. We found small but significant differences in expression of specific genes related with pluripotency and neuronal lineage commitment associated with ICSD. By reducing ICSD, it may be possible to improve yield from less start material.