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Sci Rep ; 9(1): 13190, 2019 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-31519924

RESUMO

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.


Assuntos
Genes myc , Mucosa Olfatória/citologia , Proteínas Recombinantes/genética , Transdução Genética/métodos , Adulto , Animais , Biomarcadores/metabolismo , Linhagem Celular , Técnicas de Cocultura , Gânglios Espinais/citologia , Gentamicinas/farmacologia , Humanos , Cariotipagem , Camundongos , Neuroblastoma/patologia , Mucosa Olfatória/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Receptores de Estrogênio/genética , Proteínas Recombinantes/metabolismo , Células Receptoras Sensoriais/citologia , Tamoxifeno/análogos & derivados , Tamoxifeno/farmacologia , Transgenes
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