目的建立多能干细胞系,并鉴定其是否具有正常干细胞的特性。方法将原代皮肤标本用胶原酶消化处理,利用成纤维细胞培养基进行培养,然后感染病毒。对挑选的人成体细胞来源的iPS克隆进行扩大培养,并对克隆进行免疫荧光检测和RT-PCR检测,确定表达的分子标志,采用体外三胚层分化来鉴定其生物学特性。结果所获得的iPS细胞经鉴定具有碱性磷酸酶活性,并表达Nanog、SSEA-4、Sox2、TRA-1-60等干细胞特异标记物,可形成拟胚体并在体内外可以分化为三胚层的细胞类型。结论成功建立了iPS细胞系,为今后的研究提供了良好的细胞模型。 Objective To establish pluripotent stem cell lines and identify whether they have the characteristics of normal stem cells. Methods The primary skin specimens were digested with collagenase, cultured in fibroblast medium and then infected with virus. The iPS clones from selected human adult cells were expanded and cultured. The clones were detected by immunofluorescence and RT-PCR, and the molecular markers of expression were identified. The biological characteristics of the iPS clones were identified by in vitro somatic embryo differentiation. Results The obtained iPS cells were identified as alkaline phosphatase activity and expressed as stem cell specific markers such as Nanog, SSEA-4, Sox2 and TRA-1-60, which could form embryoid bodies and differentiate into three germ layers in vivo and in vitro Of the cell type. Conclusion The successful establishment of iPS cell line provides a good cell model for future research.