Fan Yong, Luo Yumei, Chen Xinjie, Li Qing, Sun Xiaofang
Key Laboratory for Major Obstetric Diseases of Guangdong Province,The Third Affiliated Hospital of Guangzhou Medical University, Guangdong, China.
J Reprod Dev. 2012;58(4):404-9. doi: 10.1262/jrd.2011-046. Epub 2012 Apr 13.
Induced pluripotent stem cells (iPSCs) derived from somatic cells of patients represent a powerful tool for biomedical research and may have a wide range of applications in cell and gene therapy. However, the safety issues and the low efficiency associated with generating human iPSCs have limited their usage in clinical settings. The cell type used to create iPSCs can significantly influence the reprogramming efficiency and kinetics. Here, we show that amniotic fluid cells from the prenatal diagnosis of a β-thalassemia patient can be efficiently reprogrammed using a doxycycline (DOX)-inducible humanized version of the single lentiviral "stem cell cassette" vector flanked by loxP sites, which can be excised with Cre recombinase. We also demonstrated that the patient-derived iPSCs can be characterized based on the expression of pluripotency markers, and they can be differentiated into various somatic cell types in vitro and in vivo. Moreover, microarray analysis demonstrates a high correlation coefficient between human β-thalassemia iPS cells and human embryonic stem (hES) cells but a low correlation coefficient between human β-thalassemia amniotic fluid cells and human β-thalassemia iPS cells. Our data suggest that amniotic fluid cells may be an ideal human somatic cell resource for rapid and efficient generation of patient-specific iPS cells.
源自患者体细胞的诱导多能干细胞（iPSC）是生物医学研究的有力工具，在细胞和基因治疗中可能有广泛应用。然而，与生成人类iPSC相关的安全问题和低效率限制了它们在临床环境中的使用。用于创建iPSC的细胞类型可显著影响重编程效率和动力学。在此，我们表明，使用由loxP位点侧翼的强力霉素（DOX）诱导的人源化单慢病毒“干细胞盒”载体，可高效重编程来自一名β地中海贫血患者产前诊断的羊水细胞，该载体可用Cre重组酶切除。我们还证明，源自患者的iPSC可根据多能性标志物的表达进行表征，并且它们可在体外和体内分化为各种体细胞类型。此外，微阵列分析表明，人类β地中海贫血iPS细胞与人类胚胎干细胞（hES）之间的相关系数高，但人类β地中海贫血羊水细胞与人类β地中海贫血iPS细胞之间的相关系数低。我们的数据表明，羊水细胞可能是快速高效生成患者特异性iPS细胞的理想人类体细胞资源。
