Oh Jin Kyo, Przepiorski Aneta, Chang Hao-Han, Dodd Rachel C, Sander Veronika, Sorrenson Brie, Shih Jen-Hsing, Hollywood Jennifer A, de Zoysa Janak R, Shepherd Peter R, Davidson Alan J, Holm Teresa M
Department of Molecular Medicine and Pathology, University of Auckland, Auckland, New Zealand.
Maurice Wilkins Centre, Auckland, New Zealand.
J R Soc N Z. 2020 Oct 18;52(1):54-67. doi: 10.1080/03036758.2020.1830808. eCollection 2022.
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 layers and 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.
我们旨在从新西兰捐赠者中生成人类诱导多能干细胞(iPSC)系。这些细胞系是在新西兰首次生成的。从两名个体捐赠者处收集人皮肤成纤维细胞,并使用仙台病毒系统用人OSKM转录因子进行重编程。挑选出新兴的iPSC集落,进行扩增并进行核型分析。对核型正常的克隆进行多能性标志物表达、p53突变状态和三系分化潜能的鉴定。MANZ-2-2和MANZ-4-37 iPSC系显示核型正常,在RNA和蛋白质水平表达多能性标志物,且未检测到转基因表达。这两个细胞系均分化为三个胚层,并通过了hPSC多能性和三系分化计分卡检测。此外,这两个细胞系对nutlin-3a介导的细胞凋亡敏感,表明其p53野生型状态。本研究展示了从新西兰捐赠者中成功获得并鉴定的iPSC系。这些细胞系将促进新西兰及其他地区基于iPSC的研究。