Pluripotency can be rapidly and efficiently induced in human amniotic fluid-derived cells
Pluripotency can be rapidly and efficiently induced in human amniotic fluid-derived cells
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DOI：
10.1093/hmg/ddp386
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发表时间：
2009-11-15
影响因子：
3.5
通讯作者：
Jin, Ying
中科院分区：
文献类型：
--
作者：
Li, Chunliang;Zhou, Junmei;Jin, Ying
将人类体细胞直接重编程为多能性在产生用于疾病建模和细胞替代疗法的患者特异性诱导多能干（iPS）细胞方面具有广泛的意义。然而，与产生人iPS细胞相关的低效率和安全性问题限制了它们在临床环境中的使用。细胞类型可显著影响重编程效率和动力学。迄今为止，人类iPS细胞仅从少数细胞类型获得。在这里，我们首次报道了通过异位表达四种人类因子：OCT 4/SOX 2/KLF 4/C-MYC，从人羊水来源的细胞（hAFDCs）快速有效地产生iPS细胞。值得注意的是，典型的单个iPS细胞集落可以在病毒感染后6天以高效率拾取。已经衍生了八种iPS细胞系。它们可以在体外连续繁殖，并表达多能性标记物，如AKP、OCT 4、SOX 2、SSEA 4、TRA-1-60和TRA 1 -81，保持正常核型。在建立的iPS细胞系中，转基因完全失活，内源性OCT 4启动子充分去甲基化。此外，在拟胚体和畸胎瘤中分别发现了来自所有三个胚层的各种细胞和组织。此外，微阵列分析显示hAFDC-iPS细胞与人胚胎干细胞之间的相关系数高，但hAFDC-iPS细胞与hAFDC-iPS细胞之间的相关系数低。总之，这些数据确定了一个理想的人类体细胞资源，用于快速有效地产生iPS细胞，使我们能够使用更先进的方法建立人类iPS细胞，并可能建立疾病或患者特异性iPS细胞。
Direct reprogramming of human somatic cells into pluripotency has broad implications in generating patient-specific induced pluripotent stem (iPS) cells for disease modeling and cellular replacement therapies. However, the low efficiency and safety issues associated with generation of human iPS cells have limited their usage in clinical settings. Cell types can significantly influence reprogramming efficiency and kinetics. To date, human iPS cells have been obtained only from a few cell types. Here, we report for the first time rapid and efficient generation of iPS cells from human amniotic fluid-derived cells (hAFDCs) via ectopic expression of four human factors: OCT4/SOX2/KLF4/C-MYC. Significantly, typical single iPS cell colonies can be picked up 6 days after viral infection with high efficiency. Eight iPS cell lines have been derived. They can be continuously propagated in vitro and express pluripotency markers such as AKP, OCT4, SOX2, SSEA4, TRA-1-60 and TRA1-81, maintaining the normal karyotype. Transgenes are completely inactivated and the endogenous OCT4 promoter is adequately demethylated in the established iPS cell lines. Moreover, various cells and tissues from all three germ layers are found in embryoid bodies and teratomas, respectively. In addition, microarray analysis demonstrates a high correlation coefficient between hAFDC-iPS cells and human embryonic stem cells, but a low correlation coefficient between hAFDCs and hAFDC-iPS cells. Taken together, these data identify an ideal human somatic cell resource for rapid and efficient generation of iPS cells, allowing us to establish human iPS cells using more advanced approaches and possibly to establish disease-or patient-specific iPS cells.