Serum Starvation Induced Cell Cycle Synchronization Facilitates Human Somatic Cells Reprogramming
Serum Starvation Induced Cell Cycle Synchronization Facilitates Human Somatic Cells Reprogramming
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血清饥饿诱导的细胞周期同步促进人类体细胞重编程
DOI：
10.1371/journal.pone.0028203
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发表时间：
2012-04-18
期刊：
影响因子：
3.7
通讯作者：
Ge, Jian
中科院分区：
文献类型：
--
作者：
Chen, Mengfei;Huang, Jingjing;Ge, Jian
人类诱导多能干细胞（iPSC）为再生医学和人类疾病研究提供了一个有价值的模型。然而，迄今为止，人类成体细胞的重编程效率仍然很低。最近的研究表明，细胞周期是驱动表观遗传重编程多能性的关键参数。众所周知，逆转录病毒如莫洛尼鼠白血病病毒（MoMLV）需要细胞分裂以整合到宿主基因组中并复制，而用于重编程的靶原代细胞是具有不同细胞周期节律的几种细胞类型的混合物。细胞周期同步化是否对逆转录病毒诱导的重编程有潜在的影响还没有详细说明。在这项研究中，利用瞬时血清饥饿诱导的同步化，我们证明了饥饿产生可逆的细胞周期停滞，并在释放后同步进行G2/M期，大大提高了逆转录病毒感染效率。有趣的是，同步化的人皮肤成纤维细胞（HDF）和脂肪干细胞（ASC）在感染后表现出比正常FBS对照更均一的上皮形态，并且上皮标志物如E-cadherin和Epcam的表达被强烈激活。同步化处理最终提高了Nanog阳性克隆数，达到了15-20倍的增加。这些结果表明，细胞周期同步促进间充质上皮转化（MET），并促进逆转录病毒介导的重编程。我们的研究，利用血清饥饿，而不是额外的化学品，提供了一个新的见解细胞周期调控和诱导重编程的人类细胞。
Human induced pluripotent stem cells (iPSCs) provide a valuable model for regenerative medicine and human disease research. To date, however, the reprogramming efficiency of human adult cells is still low. Recent studies have revealed that cell cycle is a key parameter driving epigenetic reprogramming to pluripotency. As is well known, retroviruses such as the Moloney murine leukemia virus (MoMLV) require cell division to integrate into the host genome and replicate, whereas the target primary cells for reprogramming are a mixture of several cell types with different cell cycle rhythms. Whether cell cycle synchronization has potential effect on retrovirus induced reprogramming has not been detailed. In this study, utilizing transient serum starvation induced synchronization, we demonstrated that starvation generated a reversible cell cycle arrest and synchronously progressed through G2/M phase after release, substantially improving retroviral infection efficiency. Interestingly, synchronized human dermal fibroblasts (HDF) and adipose stem cells (ASC) exhibited more homogenous epithelial morphology than normal FBS control after infection, and the expression of epithelial markers such as E-cadherin and Epcam were strongly activated. Futhermore, synchronization treatment ultimately improved Nanog positive clones, achieved a 15–20 fold increase. These results suggested that cell cycle synchronization promotes the mesenchymal to epithelial transition (MET) and facilitates retrovirus mediated reprogramming. Our study, utilization of serum starvation rather than additional chemicals, provide a new insight into cell cycle regulation and induced reprogramming of human cells.