目的 建立WNT1基因突变(WNT1c.677C>T)所致成骨不全症患者皮肤组织来源的诱导多能干细胞(induced pluripotent stem cells，iPSCs)系，为疾病相关的分子机制研究和干细胞治疗提供新的细胞模型。 方法 通过Sanger测序明确患者的致病变异。在征得患者知情同意后，采集其皮肤组织样本，原代培养皮肤成纤维细胞。利用仙台病毒介导的非基因组整合的体细胞重编程方法，将皮肤成纤维细胞诱导为iPSCs，并对其进行多能性、分化能力以及核型鉴定。 结果 患者携带WNT1基因c.677C>T(p.Ser226Leu)纯合错义突变。由其皮肤成纤维细胞重编程建立的iPSCs系具备自我更新及体外分化能力，核型为正常的二倍体(46，XX)。 结论 建立了患者来源的WNT1基因突变(WNT1c.677C>T)iPSCs系，为该突变所致成骨不全症的研究提供了新的细胞模型。 Objective To obtain skin-derived induced pluripotent stem cells (iPSCs) from an Osteogenesis imperfecta (OI) patient carrying WNT1c.677C>T mutation in order to provide a new cell model for investigating the underlying molecular mechanism and stem cell therapy for OI. Methods The pathogenic variant of the patient was identified by Sanger sequencing. With informed consent from the patient, skin tissue was biopsied, and primary skin fibroblasts were cultured. Skin fibroblasts were induced into iPSCs using Sendai virus-mediated non-genomic integration reprogramming method. The iPSC cell lines were characterized for pluripotency, differentiation capacity, and karyotyping assay. Results The patient was found to carry homozygous missense c. 677C>T (p.Ser226Leu) mutation of theWNT1 gene. The established iPSC lines possessed self-renewal and capacity for in vitro differentiation. It also has a diploid karyotype (46, XX). Conclusion A patient-specific WNT1 gene mutation (WNT1c.677C>T) iPSC line was established, which can provide a cell model for the study of OI caused by the mutation.
Generation of skin-derived iPSCs from an Osteogenesis imperfecta patient carryingWNT1 c.677C>T mutation
Objective To obtain skin-derived induced pluripotent stem cells (iPSCs) from an Osteogenesis imperfecta (OI) patient carrying WNT1c.677C>T mutation in order to provide a new cell model for investigating the underlying molecular mechanism and stem cell therapy for OI. Methods The pathogenic variant of the patient was identified by Sanger sequencing. With informed consent from the patient, skin tissue was biopsied, and primary skin fibroblasts were cultured. Skin fibroblasts were induced into iPSCs using Sendai virus-mediated non-genomic integration reprogramming method. The iPSC cell lines were characterized for pluripotency, differentiation capacity, and karyotyping assay. Results The patient was found to carry homozygous missense c. 677C>T (p.Ser226Leu) mutation of theWNT1 gene. The established iPSC lines possessed self-renewal and capacity for in vitro differentiation. It also has a diploid karyotype (46, XX). Conclusion A patient-specific WNT1 gene mutation (WNT1c.677C>T) iPSC line was established, which can provide a cell model for the study of OI caused by the mutation.
Osteogenesis imperfectaWNT1 c. 677C>T mutationInduced pluripotent stem cellSendai virus
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