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利用转座子载体将大鼠胚胎成纤维细胞重编程为诱导多能干细胞及其体外软骨分化

Reprogramming rat embryonic fibroblasts into induced pluripotent stem cells using transposon vectors and their chondrogenic differentiation in vitro.

作者信息

Ye Jiakuan, Hong Junyi, Ye Fusheng

机构信息

Department of Orthopaedics, Xiaoshan Hospital, Hangzhou, Zhejiang 311202, P.R. China.

出版信息

Mol Med Rep. 2015 Feb;11(2):989-94. doi: 10.3892/mmr.2014.2793. Epub 2014 Oct 27.

Abstract

It has been known that the successful reprogramming of differentiated human somatic cells into a pluripotent state would allow for the creation of cartilage cells. However, current virus‑mediated strategies to form induced pluripotent stem cells (iPSCs) are limited in their clinical application due to exogenous gene modification. In the present study, the piggyBac transposon system carrying corresponding genes (Oct3/4, Klf4, Sox2 and c‑Myc) was employed to reprogram rat embryonic fibroblasts (REFs) into iPSCs, and the transposon‑carried genes were successfully removed by a transposase system. Furthermore, the differentiation capacity of the iPSCs into chondrocytes was investigated in vitro. A typical chondrocyte marker, collagen‑II, was expressed following culture. In conclusion, rat iPSCs without genetic modification were obtained from REFs using the piggyBac transposon system and the transposase mPB system, and these cells possessed the capacity of chondrocyte differentiation, suggesting that this method may be an effective approach for the treatment of cartilage disorders in the future.

摘要

已知将分化的人类体细胞成功重编程为多能状态可用于软骨细胞的生成。然而,当前通过病毒介导形成诱导多能干细胞(iPSC)的策略由于外源基因修饰而在临床应用中受到限制。在本研究中,利用携带相应基因(Oct3/4、Klf4、Sox2和c-Myc)的piggyBac转座子系统将大鼠胚胎成纤维细胞(REF)重编程为iPSC,并且通过转座酶系统成功去除了转座子携带的基因。此外,在体外研究了iPSC向软骨细胞的分化能力。培养后表达了典型的软骨细胞标志物II型胶原。总之,使用piggyBac转座子系统和转座酶mPB系统从REF获得了无基因修饰的大鼠iPSC,并且这些细胞具有软骨细胞分化能力,表明该方法可能是未来治疗软骨疾病的有效途径。

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