Neuron-like differentiation and selective ablation of undifferentiated embryonic stem cells containing suicide gene with Oct-4 promoter
Neuron-like differentiation and selective ablation of undifferentiated embryonic stem cells containing suicide gene with Oct-4 promoter
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DOI：
10.1089/scd.2007.0235
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发表时间：
2008-08-01
影响因子：
4
通讯作者：
Mori, Hideki
中科院分区：
文献类型：
--
作者：
Hara, Akira;Aoki, Hitomi;Mori, Hideki
未分化的胚胎干细胞（ES细胞）体内移植可产生畸胎瘤，细胞增殖不受控制。虽然ES细胞可能是未来人类细胞替代疗法的有吸引力的候选者，但其应用于治疗的主要限制是畸胎瘤的形成。在本研究中，ES细胞含有单纯疱疹病毒-胸苷激酶（HSV-tk）转基因的自杀基因表达的控制下的Oct-4启动子被用于消融未分化的ES细胞，这可能会产生畸胎瘤，使用三维细胞培养系统，允许一个多层的细胞结构。用更昔洛韦处理未分化的ES细胞，实现了Oct-4启动子控制下表达HSV-tk基因的选择性消融。更昔洛韦治疗后存活的ES细胞表达了几种神经元相关的标志物，如突触素，β-微管蛋白，囊泡谷氨酸转运蛋白1，突触融合蛋白，蛋白激酶C和胶质细胞酸性蛋白（GFAP），但不是Oct-4。在存活的ES细胞中，作为神经元突触标志物的突触素和作为胶质纤维标志物的GFAP的共表达揭示了精细结构的神经元网络。存活的ES细胞Ki-67增殖指数下降。总之，自杀基因对未分化ES细胞的选择性消融降低了ES细胞的增殖活性并诱导了ES细胞的神经元样分化。
In vivo transplantation of undifferentiated embryonic stem (ES) cells can produce teratomas with uncontrolled cell proliferation. Although ES cells may be attractive candidates for human cell-replacement therapy in the future, the major limitation of its application to the therapy is teratoma formation. In the present study, ES cells containing herpes simplex virus-thymidine kinase (HSV-tk) transgene for a suicide gene expression under the control of the Oct-4 promoter was used for ablation of undifferentiated ES cells, which may produce teratomas, using three-dimensional cell culture system allowing a multilayer cell construct. Selective ablation of undifferentiated ES cells expressing HSV-tk gene under the control of Oct-4 promoter was achieved by ganciclovir treatment. Surviving ES cells after ganciclovir treatment expressed several neuron-associated markers such as synaptophysin, beta-tubulin, vesicular glutamate transporter 1, syntaxin, protein kinace C and glial fibrillary acidic protein (GFAP) but not Oct-4. Coexpression of synaptophysin as a marker of neuronal synapse and GFAP as that of glial fibers in the surviving ES cells revealed finely structured neuronal network. Furthermore, decrease of Ki-67 proliferative index was detected in the surviving ES cells. In conclusion, selective ablation of undifferentiated ES cells by a suicide gene decreases proliferative activity and induces neuron-like differentiation in ES cells.