Towards The Development Of An Integrated Scalable Bioprocess For The Production Of HiPSC-Derived NPC

Abstract

Neural precursor cells (NPCs) can be used as a cell source for disease modelling, tissue engineering and cell-based regenerative therapies for neurological disorders. However, due to the limited availability of NPCs sourced from foetal tissues, ethical concerns and invasive methods, their use in late-stage clinical trials and commercial availability is hindered. Human induced pluripotent stem cells (hiPSCs) have the potential to differentiate towards NPCs, thus providing an alternative unlimited source for generating NPCs which bypasses the current sourcing limitations. hiPSC-derived NPCs have been successfully produced using direct or indirect methods. However, those methods have predominantly been carried out using traditional planar culture systems. To achieve the high cell numbers required for therapeutic treatments, translation of these protocols to scalable production methods is necessary. In this thesis, we have developed an integrated bioprocess for production of hiPSC-derived NPCs. This would enable the development of a process using dynamic 3D suspended culture systems and stirred tank bioreactors (STBs), which are scalable and relevant for clinical productions of hiPSC-derived NPCs. We have identified a suitable microcarrier for hiPSC expansion by performing systematic screenings of 8 commercially available microcarriers. We have also assessed the potential of microcarriers as a platform for the neural induction of hiPSC. We have deployed these microcarriers to develop towards a scalable integrated bioprocess in spinner flasks. Overall, we demonstrated the translation of the baseline and microcarrier integrated bioprocesses towards a scalable bioprocess utilising scalable spinner flasks. However, although spinner flasks have been successfully employed for the expansion of F.D., de la Raga, PhD Thesis, Aston University, 2022 3 mammalian cells such as Chinese hamster ovary (CHO) or human embryonic kidney (HEK) cell lines, results here show that the expansion and neuralisation of hiPSCs on tissue culture plastic planar platforms remained superior in terms of maintaining pluripotency, growth kinetics and differentiation potential.