Generation of c-MycER<sup>TAM</sup>-transduced human late-adherent olfactory mucosa cells for potential regenerative applications

Type of content
Journal Article
Thesis discipline
Degree name
Publisher
Springer Science and Business Media LLC
Journal Title
Journal ISSN
Volume Title
Language
eng
Date
2019
Authors
Santiago-Toledo , Gerardo
Georgiou , Melanie
dos Reis , Joana
Roberton , Victoria H.
Valinhas , Ana
Wood, Rachael
Phillips , James B.
Mason , Chris
Li , Daqing
Li , Ying
Abstract

Human olfactory mucosa cells (hOMCs) have been transplanted to the damaged spinal cord both pre-clinically and clinically. To date mainly autologous cells have been tested. However, inter-patient variability in cell recovery and quality, and the fact that the neuroprotective olfactory ensheathing cell (OEC) subset is difficult to isolate, means an allogeneic hOMC therapy would be an attractive “off-the-shelf” alternative. The aim of this study was to generate a candidate cell line from late-adherent hOMCs, thought to contain the OEC subset. Primary late-adherent hOMCs were transduced with a c-MycERTAM gene that enables cell proliferation in the presence of 4-hydroxytamoxifen (4-OHT). Two c-MycERTAM-derived polyclonal populations, PA5 and PA7, were generated and expanded. PA5 cells had a normal human karyotype (46, XY) and exhibited faster growth kinetics than PA7, and were therefore selected for further characterisation. PA5 hOMCs express glial markers (p75NTR, S100ß, GFAP and oligodendrocyte marker O4), neuronal markers (nestin and ß-III-tubulin) and fibroblast-associated markers (CD90/Thy1 and fibronectin). Co-culture of PA5 cells with a neuronal cell line (NG108-15) and with primary dorsal root ganglion (DRG) neurons resulted in significant neurite outgrowth after 5 days. Therefore, c-MycERTAM-derived PA5 hOMCs have potential as a regenerative therapy for neural cells.

Description
Citation
Santiago-Toledo G, Georgiou M, dos Reis J, Roberton VH, Valinhas A, Wood RC, Phillips JB, Mason C, Li D, Li Y, Sinden JD, Choi D, Jat PS, Wall IB (2019). Generation of c-MycER<sup>TAM</sup>-transduced human late-adherent olfactory mucosa cells for potential regenerative applications. Scientific Reports. 9(1). 13190-.
Keywords
Olfactory Mucosa, Ganglia, Spinal, Cell Line, Rats, Sprague-Dawley, Neuroblastoma, Tamoxifen, Gentamicins, Receptors, Estrogen, Recombinant Proteins, Coculture Techniques, Karyotyping, Transduction, Genetic, Genes, myc, Transgenes, Sensory Receptor Cells, Biomarkers
Ngā upoko tukutuku/Māori subject headings
ANZSRC fields of research
32 - Biomedical and clinical sciences::3209 - Neurosciences::320902 - Cellular nervous system
32 - Biomedical and clinical sciences::3209 - Neurosciences::320905 - Neurology and neuromuscular diseases
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All rights reserved unless otherwise stated