Baharvand Hossein, Totonchi Mehdi, Taei Adeleh, Seifinejad Ali, Aghdami Nasser, Salekdeh Ghasem Hosseini
Department of Stem Cells, Cell Science Research Center, Royan Institute, ACECR, Tehran, Iran.
Methods Mol Biol. 2010;584:425-43. doi: 10.1007/978-1-60761-369-5_23.
The recent discovery of genomic reprogramming of human somatic cells to an embryonic stem (ES) cell-like pluripotent state provides a unique opportunity for stem cell research. The reprogrammed cells, named as induced pluripotent stem (iPS) cells, possess many of the properties of ES cells and represent one of the most promising sources of patient-specific cells for use in disease model, development of pharmacology and toxicology, screening teratogens, and regenerative medicine. Here we describe the detailed methods for the generation of undifferentiated human iPS (hiPS) cells in feeder layer- and serum-free conditions. This system eliminates direct contact of stem cells with MEFs and reduces use of unknown serum factors that may have undesired activities and enables consistency in large-scale and long-term expansion of undifferentiated hiPS cells. Our findings greatly simplify the method for induction of pluripotency and bring it one step closer to clinical applications. Moreover, the established hiPS cells showed chromosomal stability during long-term culture.
最近人类体细胞基因组重编程为胚胎干细胞（ES）样多能状态的发现为干细胞研究提供了独特的机会。这些重编程细胞被命名为诱导多能干细胞（iPS细胞），具有许多ES细胞的特性，是用于疾病模型、药理学和毒理学发展、致畸剂筛选以及再生医学的最有前景的患者特异性细胞来源之一。在此，我们描述了在无饲养层和无血清条件下生成未分化人类iPS（hiPS）细胞的详细方法。该系统消除了干细胞与小鼠胚胎成纤维细胞（MEFs）的直接接触，并减少了可能具有不良活性的未知血清因子的使用，且能使未分化hiPS细胞在大规模和长期传代培养中保持一致性。我们的发现极大地简化了多能性诱导方法，并使其更接近临床应用。此外，所建立的hiPS细胞在长期培养过程中显示出染色体稳定性。
