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首页> 外文期刊>Journal of Cellular Physiology >The influence of rAAV2‐mediated SOX2 delivery into neonatal and adult human RPE cells; a comparative study
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The influence of rAAV2‐mediated SOX2 delivery into neonatal and adult human RPE cells; a comparative study

机译:rAAV2介导SOX2应承担交付的影响新生儿和成人RPE细胞;研究

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Cell replacement is a promising therapy for degenerative diseases like age‐related macular degeneration (AMD). Since the human retina lacks regeneration capacity, much attention has been directed toward persuading for cells that can differentiate into retinal neurons. In this report, we have investigated reprogramming of the human RPE cells and concerned the effect of donor age on the cellular fate as a critical determinant in reprogramming competence. We evaluated the effect of SOX2 over‐expression in human neonatal and adult RPE cells in cultures. The coding region of human SOX2 gene was cloned into adeno‐associated virus (AAV2) and primary culture of human neonatal/adult RPE cells were infected by recombinant virus. De‐differentiation of RPE to neural/retinal progenitor cells was investigated by quantitative real‐time PCR and ICC for neural/retinal progenitor cells’ markers. Gene expression analysis showed 80‐fold and 12‐fold over‐expression for SOX2 gene in infected neonatal and adult hRPE cells, respectively. The fold of increase for Nestin in neonatal and adult hRPE cells was 3.8‐fold and 2.5‐fold, respectively. PAX6 expression was increased threefold and 2.5‐fold in neonatal/adult treated cultures. Howbeit, we could not detect rhodopsin, and CHX10 expression in neonatal hRPE cultures and expression of rhodopsin in adult hRPE cells. Results showed SOX2 induced human neonatal/adult RPE cells to de‐differentiate toward retinal progenitor cells. However, the increased number of PAX6, CHX10, Thy1, and rhodopsin positive cells in adult hRPE treated cultures clearly indicated the considerable generation of neuro‐retinal terminally differentiated cells.
机译:细胞替代是一个有前途的治疗退行性疾病如年龄相关的黄斑变性(AMD)。再生能力,关注为了说服细胞分化成视网膜神经元。报告中,我们调查的重组有关人类RPE细胞和捐赠的效果年龄对细胞命运的关键行列式在重编程能力。评估SOX2在量表达的影响人类新生儿和成人文化RPE细胞。克隆人类SOX2基因的编码区在腺病毒相关病毒(AAV2)和主文化的人类新生儿/成人RPE细胞被重组病毒感染。RPE神经/视网膜祖细胞通过定量实时PCR和调查ICC神经/视网膜祖细胞的标记。基因表达分析显示80折12公/褶皱量表达式SOX2在感染的基因新生儿和成人hRPE细胞,分别。褶皱巢蛋白的增加新生儿和成人hRPE细胞是3.8折,2.5折,应承担的分别。三倍和2.5折在新生儿/成人治疗文化。与新生儿hRPE CHX10表达文化和表达的视紫红质在成人hRPE细胞。结果显示SOX2诱导人类新生儿/成人RPE细胞向视网膜de应承担的区分祖细胞。PAX6、CHX10 Thy1,视紫红质阳性成人hRPE细胞治疗文化明显显示相当大的一代神经视网膜终末分化细胞。

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