title: Isolation of Live Premature Senescent Cells Using FUCCI Technology.
abstract: Cellular senescence plays an important role in diverse biological processes such as tumorigenesis and organismal aging. However, lack of methods to specifically identify and isolate live senescent cells hampers the precise understanding of the molecular mechanisms regulating cellular senescence. Here, we report that utilization of fluorescent ubiquitination-based cell cycle indicator (FUCCI) technology allows isolation of live premature senescent cells induced by doxorubicin treatment. Exposure of human foreskin fibroblasts (HFFs) to a low dose of doxorubicin led to cellular senescent phenotypes including formation of γ-H2AX and 53BP1 foci indicative of DNA damage, decreased cell proliferation and increased senescence-associated β-galactosidase (SA-β-gal) activity. Importantly, doxorubicin-induced senescent cells were arrested at S/G2/M phases of cell cycle which can be reported by a construct encoding a fragment of hGeminin fused with monomeric Azami-Green (mAG-hGeminin). Flow cytometric sorting of GFP(+) cells from doxorubicin-treated HFFs carrying mAG-hGeminin reporter enabled isolation and enrichment of live senescent cells in the culture. Our study develops a novel method to identify and isolate live premature senescent cells, thereby providing a new tool to study cellular senescence.
authors: Dr. Li Dak Sum &Yip Yio Chin Center for Stem Cell and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China.|Department of Pathophysiology, Anhui Medical University, Hefei 230032, China.
authors: Danli
authors: D
authors: Wang
authors: Dr. Li Dak Sum &Yip Yio Chin Center for Stem Cell and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China.
authors: Ping
authors: P
authors: Lu
authors: Dr. Li Dak Sum &Yip Yio Chin Center for Stem Cell and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China.
authors: Yang
authors: Y
authors: Liu
authors: Dr. Li Dak Sum &Yip Yio Chin Center for Stem Cell and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China.
authors: Li
authors: L
authors: Chen
authors: Dr. Li Dak Sum &Yip Yio Chin Center for Stem Cell and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China.
authors: Rui
authors: R
authors: Zhang
authors: Dr. Li Dak Sum &Yip Yio Chin Center for Stem Cell and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China.
authors: Weihao
authors: W
authors: Sui
authors: Dr. Li Dak Sum &Yip Yio Chin Center for Stem Cell and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China.
authors: Alexandru George
authors: AG
authors: Dumitru
authors: Division of Hematology and Oncology, Shenzhen Children's Hospital, 7019 Yitian Road, Futian District, Shenzhen, China.
authors: Xiaowen
authors: X
authors: Chen
authors: Division of Hematology and Oncology, Shenzhen Children's Hospital, 7019 Yitian Road, Futian District, Shenzhen, China.
authors: Feiqiu
authors: F
authors: Wen
authors: Dr. Li Dak Sum &Yip Yio Chin Center for Stem Cell and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China.|Zhejiang Provincial Key Laboratory of Tissue Engineering and Regenerative Medicine, Hangzhou, China.
authors: Hong-Wei
authors: HW
authors: Ouyang
authors: Dr. Li Dak Sum &Yip Yio Chin Center for Stem Cell and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China.|Zhejiang Provincial Key Laboratory of Tissue Engineering and Regenerative Medicine, Hangzhou, China.
authors: Junfeng
authors: J
authors: Ji
mesh_terms: D059447:Cell Cycle Checkpoints / Q000187:drug effects*; D002469:Cell Separation / Q000295:instrumentation* / Q000379:methods*; D016922:Cellular Senescence / Q000187:drug effects*; D004249:DNA Damage; D004317:Doxorubicin / Q000494:pharmacology*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D006801:Humans
chemical_list: D004317:Doxorubicin