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Serum starvation induced cell cycle synchronization facilitates human somatic cells reprogramming

Chen, M and Huang, J and Yang, X and Liu, B and Zhang, W and Huang, L and Deng, F and Ma, J and Bai, Y and Lu, R and Huang, B and Gao, Q and Zhuo, Y and Ge, J (2012) Serum starvation induced cell cycle synchronization facilitates human somatic cells reprogramming. PLoS ONE, 7 (4).

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Abstract

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. © 2012 Chen et al.

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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Chen, M
Huang, J
Yang, X
Liu, B
Zhang, W
Huang, L
Deng, F
Ma, J
Bai, Y
Lu, R
Huang, B
Gao, Q
Zhuo, Y
Ge, J
Contributors:
ContributionContributors NameEmailPitt UsernameORCID
EditorAllodi, SilvanaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Date: 18 April 2012
Date Type: Publication
Journal or Publication Title: PLoS ONE
Volume: 7
Number: 4
DOI or Unique Handle: 10.1371/journal.pone.0028203
Schools and Programs: School of Medicine > Microbiology and Molecular Genetics
Refereed: Yes
Other ID: NLM PMC3329488
PubMed Central ID: PMC3329488
PubMed ID: 22529890
Date Deposited: 24 Sep 2012 20:26
Last Modified: 02 Feb 2019 16:56
URI: http://d-scholarship.pitt.edu/id/eprint/14173

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