Devito Liani, Donne Matthew, Kolundzic Nikola, Khurana Preeti, Hobbs Carl, Kaddour Gabriel, Dubrac Sandrine, Gruber Robert, Schmuth Matthias, Mauro Thea, Ilic Dusko
Stem Cell Laboratory, Department of Women and Children's Health, School of Life Course Sciences, Faculty of Life Sciences and Medicine, King's College London, London, UK; Assisted Conception Unit, Guy's Hospital, London, UK.
VitroLabs Inc., San Francisco, California, USA.
Stem Cell Res. 2018 Aug;31:122-126. doi: 10.1016/j.scr.2018.07.014. Epub 2018 Jul 25.
We have generated an induced pluripotent stem cell (iPSC) line KCLi001-A (iOP118) from a female atopic dermatitis (AD) patient, heterozygous for the loss-of-function mutation c.2282del4 in the filaggrin gene (FLG). Epidermal keratinocytes were reprogrammed using non-integrating Sendai virus vectors. The entire process of derivation and expansion of AD-iPSCs were performed under xeno-free culture conditions. Characterization of KCLi001-A line included molecular karyotyping, mutation screening using restriction enzyme digestion and Sanger sequencing, while pluripotency and differentiation potential were confirmed by expression of associated markers in vitro and by in vivo teratoma assay.
我们从一名患有特应性皮炎(AD)的女性患者中生成了诱导多能干细胞(iPSC)系KCLi001-A(iOP118),该患者的丝聚合蛋白基因(FLG)存在功能丧失突变c.2282del4的杂合子。使用非整合型仙台病毒载体对表皮角质形成细胞进行重编程。AD-iPSC的整个衍生和扩增过程均在无血清培养条件下进行。KCLi001-A系的鉴定包括分子核型分析、使用限制性酶切和桑格测序进行突变筛查,而多能性和分化潜能则通过体外相关标志物的表达及体内畸胎瘤试验得以证实。