Objective

Pluripotent stem cells carry the innate ability to proliferate and differentiate into any cell type and provide a unique opportunity for improving medical treatment and knowledge through disease models, replacement therapy and drug screening. Huge strides have been made in improving the efficiency and purity of stem cell maintenance and differentiation protocols; however most protocols still use conventional culture plates which are challenging to scale up and lack complexity. We have developed a robust platform for expanding and differentiating Human Embryonic Stem cells (hES) in electrospun 3D scaffolds manufactured from biodegradable and biocompatible compounds and be used inside proprietary perfusion bioreactor systems. This platform is flexible with parameters such as volume, fibre diameter, porosity and bioactive coatings. In this study hES cells are cultured in electrospun membrane under static conditions and tested for pluripotency by staining for Nanog and Oct3/4 markers at various time intervals. Selected experiments were taken forward to undergo a neural progeny differentiation by applying a dual SMAD inhibition while still within the electrospun scaffold. RNA and immunocytochemistry analysis demonstrated the successful differentiation of the stem cell population into mature neural lineages within 21 days.

This project was supported by FP7-Health-2013-601700 (HESUB).