Generation of c-MycERTAM-transduced human late-adherent olfactory mucosa cells for potential regenerative applications.
Generation of c-MycERTAM-transduced human late-adherent olfactory mucosa cells for potential regenerative applications.
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生成 c-MycERTAM 转导的人类晚期贴壁嗅粘膜细胞，用于潜在的再生应用。
DOI：
10.1038/s41598-019-49315-6
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发表时间：
2019
影响因子：
4.6
通讯作者：
Santiago-Toledo G
中科院分区：
文献类型：
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作者：
Santiago-Toledo G
人类嗅觉粘膜细胞（hOMCs）在临床前和临床中都被移植到受损的脊髓中。迄今为止，主要是对自体细胞进行了测试。然而，患者间细胞恢复和质量的差异，以及神经保护性嗅鞘细胞（OEC）亚群难以分离的事实，意味着同种异体hOMC治疗将是一种有吸引力的“现成”替代方案。本研究的目的是从被认为包含OEC亚群的晚贴壁hOMCs中产生候选细胞系。原发晚期贴壁hOMCs被c-MycERTAMgene转导，该基因能够在4-羟基他莫昔芬（4-OHT）存在下使细胞增殖。对两个c- myceram衍生的多克隆群体PA5和PA7进行扩增。PA5细胞具有正常的人类核型（46,xy），并且比PA7表现出更快的生长动力学，因此被选中进行进一步的表征。PA5 hOMCs表达胶质标记物（p75NTR、S100ß、GFAP和少突胶质细胞标记物O4）、神经元标记物（巢蛋白和ß- iii -微管蛋白）和成纤维细胞相关标记物（CD90/Thy1和纤维连接蛋白）。PA5细胞与神经细胞系（NG108-15）和初级背根神经节（DRG）神经元共培养5天后，神经突明显生长。因此，c- myceram衍生的PA5 hOMCs具有神经细胞再生治疗的潜力。
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-MycERTAMgene 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.