人端粒酶反转录酶重组绿色荧光表达载体转染椎间盘正常髓核细胞

Recombinant green fluorescent protein plasmids expressing human telomerase reverse transcriptase transfect normal nucleus pulposus cells in the intervertebral disc

背景:椎间盘髓核细胞分离培养困难,老化较快,迫切需要一种标准细胞株用于实验研究。目的:探讨人端粒酶反转录酶重组绿色荧光表达载体的构建及其转染正常髓核细胞构建永生化细胞的可行性研究。方法:通过目的基因克隆、真核表达质粒中目的基因序列测定、目的基因真核表达质粒的构建、转染人端粒酶反转录酶表达检测等步骤进行实验。结果与结论:构建出人端粒酶反转录酶重组绿色荧光表达载体,成功转染正常髓核细胞并在细胞中稳定表达。结果表明运用人端粒酶反转录酶转染椎间盘髓核细胞构建永生化细胞是一种可行的方法。

BACKGROUND： Nucleus pulposus cells are difficult to be isolated and cultured from the intervertebral disc, and the cells age rapidly. An urgent standard cell line is required for experimental research.OBJECTIVE： To investigate the construction of recombinant green fluorescent protein plasmid expressing human telomerase reverse transcriptase and to assess the feasibility of constructing immortalized human nucleus pulposus cell line.METHODS： Target gene cloning, target gene sequencing in eukaryotic expression plasmids, construction of eukaryotic expression plasmids expressing target genes, and transfection of human telomerase reverse transcriptase were performed in order.RESULTS AND CONCLUSION： Nucleus pulposus cells transfected with recombinant green fluorescent protein plasmid expressing human telomerase reverse transcriptase have a stable expression. It is anticipated that human telomerase reverse transcriptase can be useful for creating the immortalized nucleus pulposus cells.