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首页> 外文期刊>Journal of the Royal Society of New Zealand >Derivation of induced pluripotent stem cell lines from New Zealand donors
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Derivation of induced pluripotent stem cell lines from New Zealand donors

机译:从新西兰供体衍生出诱导多能干细胞系

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摘要

We aimed to generate human induced pluripotent stem cell (iPSC) lines from New Zealand donors. These lines are the first to be generated in New Zealand. Human dermal fibroblasts were collected from two individual donors and reprogrammed with the human OSKM transcription factors using the Sendai virus system. Emerging iPSC colonies were picked, expanded and karyotyped. Clones with normal karyotype were characterised for pluripotency marker expression, p53 mutational status and trilineage differentiation potential. The MANZ-2-2 and MANZ-4-37 iPSC lines showed normal karyotype and expressed pluripotency markers at RNA and protein levels without detectable transgene expression. Both lines differentiated into the three germ layersin vitroand passed the hPSC Scorecard assay for pluripotency and trilineage differentiation. Furthermore, both lines were susceptible to cell apoptosis mediated by nutlin-3a indicative of their wildtype p53 status. This study presents the successful derivation and characterisation of iPSC lines derived from New Zealand donors. These lines will facilitate iPSC-based research in New Zealand and beyond.
机译:我们的目标是从新西兰供体中产生人类诱导多能干细胞(iPSC)系。这些生产线是新西兰最早生成的生产线。从两个单独的供体中收集人真皮成纤维细胞,并使用仙台病毒系统用人OSKM转录因子重新编程。对新兴的 iPSC 菌落进行挑选、扩增和核型分析。具有正常核型的克隆具有多能性标志物表达、p53突变状态和三系分化潜力。MANZ-2-2 和 MANZ-4-37 iPSC 细胞系显示出正常的核型,并在 RNA 和蛋白质水平上表达多能性标志物,但未检测到转基因表达。两个品系在体外分化为三个胚层,并通过hPSC记分卡测定多能性和三系分化。此外,两个品系都容易受到 nutlin-3a 介导的细胞凋亡,这表明它们的野生型 p53 状态。本研究介绍了来自新西兰供体的 iPSC 细胞系的成功推导和表征。这些细胞系将促进新西兰及其他地区基于iPSC的研究。

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