Human embryonic stem cells (hESCs) have the potential to differentiate into all cell types of the three germ layers. However, various limitations hinder their use in the clinic, including possibilities of teratoma formation, xenogenic exposure through the use of Matrigel™ and feeder layers, along with poor attachment and expansion rates and inability to transport hESCs into an in vivo site.
This thesis has aimed to overcome the above limitations. Electrospun nanofibrous substrates from a purely synthetic FDA approved material have been developed and investigated for the novel use in the expansion of undifferentiated hESCs. Synergistic effects between the oxygen environment and nanofibre technology were revealed which demonstrated the expansion of pluripotent hESCs in physiological normoxia (2% O2) on these substrates, with retention of differentiation capacity. However, in hyperoxia (21% O2), hESCs cultured on these substrates dictated embryoid body formation. A range of polymers (PCL, PLLA and PLGA) were tested (aligned and random conformations) where the optimal polymer (PCL) was further investigated at 2% O2 at various fibre diameters to reveal its impact on hESC clonogenicity.