Generation of c-MycERTAM-transduced human late-adherent olfactory mucosa cells for potential regenerative applications

Gerardo Santiago-toledo, Melanie Georgiou, Joana Dos Reis, Victoria H. Roberton, Ana Valinhas, Rachael C. Wood, James B. Phillips, Chris Mason, Daqing Li, Ying Li, John D. Sinden, David Choi, Parmjit S. Jat, Ivan B. Wall

Research output: Contribution to journalArticle

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.
Original languageEnglish
Article number13190
JournalScientific Reports
Volume9
Issue number1
DOIs
Publication statusPublished - 13 Sep 2019

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Olfactory Mucosa
Cell- and Tissue-Based Therapy
Cell Line
Nestin
Oligodendroglia
Spinal Ganglia
Tubulin
Coculture Techniques
Karyotype
Fibronectins
Neuroglia
Spinal Cord
Fibroblasts
Cell Proliferation
Neurons

Bibliographical note

This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Santiago-toledo, Gerardo ; Georgiou, Melanie ; Dos Reis, Joana ; Roberton, Victoria H. ; Valinhas, Ana ; Wood, Rachael C. ; Phillips, James B. ; Mason, Chris ; Li, Daqing ; Li, Ying ; Sinden, John D. ; Choi, David ; Jat, Parmjit S. ; Wall, Ivan B. / Generation of c-MycERTAM-transduced human late-adherent olfactory mucosa cells for potential regenerative applications. In: Scientific Reports. 2019 ; Vol. 9, No. 1.
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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, S100{\ss}, 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.",
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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-MycERTAM-transduced human late-adherent olfactory mucosa cells for potential regenerative applications', Scientific Reports, vol. 9, no. 1, 13190. https://doi.org/10.1038/s41598-019-49315-6

Generation of c-MycERTAM-transduced human late-adherent olfactory mucosa cells for potential regenerative applications. / Santiago-toledo, Gerardo; Georgiou, Melanie; Dos Reis, Joana; Roberton, Victoria H.; Valinhas, Ana; Wood, Rachael C.; Phillips, James B.; Mason, Chris; Li, Daqing; Li, Ying; Sinden, John D.; Choi, David; Jat, Parmjit S.; Wall, Ivan B.

In: Scientific Reports, Vol. 9, No. 1, 13190, 13.09.2019.

Research output: Contribution to journalArticle

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AU - Georgiou, Melanie

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AU - Mason, Chris

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PY - 2019/9/13

Y1 - 2019/9/13

N2 - 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.

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