Towards an allogeneic therapy for neural regeneration
January 27-31, 2019
Injuries to the central nervous system (CNS) can be devastating. CNS injuries include those to the spinal cord, where there can be a complete loss of function below the point of injury. Spinal cord injuries impact up to 500,000 people worldwide every year and where function is lost, quality of life can be severely limited. Olfactory ensheathing cells (OECs) are a promising cell therapy candidate for treatment of neurologic injury as they have been shown to promote neuronal survival and facilitate regeneration of severed axons. Despite their unique properties, OECs are very challenging cells to work with since they are difficult to isolate, difficult to sustain in culture for prolonged periods and there is still controversy around how to characterize their identity and potency. Due to the inherent variability of OEC yield in biopsies and difficulties in growing these cells, producing an autologous therapy is not currently a viable option. Therefore, we sought to develop allogeniec OEC lines using a conditional immortalization tool. We undertook characterization work to ensure they expressed key putative OEC markers (such as p75NTR and S100β) and were able to support neuron extension in in vitro models. Subsequently, extensive bioprocess development was undertaken to investigate parameters such as: Cell culture conditions, e.g. effect of different culture media, initial seeding densities and microcarrier-based expansion Characterization process parameters, e.g. effect of culture time and pre-selection on identity marker expression, impact of detergents on identity marker measurement, and finally their impact in functional assays (including neuron co-culture to identify the impact of processing on neuronal support activity). A number of important insights have been gained from this work including: identity markers are transient making characterization challenging; and analytics methodologies employed themselves affect what is measurable. However, these insights are informing our future approach to creating candidate cell lines for treatment of neurologic injury.
Rachael Wood and Ivan Wall, "Towards an allogeneic therapy for neural regeneration" in "Advancing Manufacture of Cell and Gene Therapies VI", Dolores Baksh, GE Healthcare, USA Rod Rietze, Novartis, USA Ivan Wall, Aston University, United Kingdom Eds, ECI Symposium Series, (2019). https://dc.engconfintl.org/cell_gene_therapies_vi/35