title: Enhanced generation of human induced pluripotent stem cells by ectopic expression of Connexin 45.
abstract: Somatic cells can be successfully reprogrammed into pluripotent stem cells by the ectopic expression of defined transcriptional factors. However, improved efficiency and better understanding the molecular mechanism underlying reprogramming are still required. In the present study, a scrape loading/dye transfer assay showed that human induced pluripotent stem cells (hiPSCs) contained functional gap junctions partially contributed by Connexin 45 (CX45). We then found CX45 was expressed in human embryonic stem cells (hESCs) and human dermal fibroblasts (hDFs) derived hiPSCs. Then we showed that CX45 was dramatically upregulated during the reprogramming process. Most importantly, the ectopic expression of CX45 significantly enhanced the reprogramming efficiency together with the Yamanaka factors (OCT4, SOX2, KLF4, cMYC - OSKM), whereas knockdown of endogenous CX45 expression significantly blocked cellular reprogramming and reduced the efficiency. Our further study demonstrated that CX45 overexpression or knockdown modulated the cell proliferation rate which was associated with the reprogramming efficiency. In conclusion, our data highlighted the critical role of CX45 in reprogramming and may increase the cell division rate and result in an accelerated kinetics of iPSCs production.
authors.affiliation: Program of Stem Cells and Regenerative Medicine, Affiliated Guangzhou Women and Children's Hospital, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510623, China.|Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, China.|Department of Biology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, China.
authors.forename: Qiong
authors.initials: Q
authors.lastname: Ke
authors.affiliation: Department of Pharmacology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, China.|Lung Biology Laboratory, Department of Medicine, Division of Pulmonary, Allergy and Critical Care, Columbia University Medical Center, New York, New York, 10032, USA.
authors.forename: Li
authors.initials: L
authors.lastname: Li
authors.affiliation: Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Yao
authors.affiliation: Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, China.
authors.forename: Xingqiang
authors.initials: X
authors.lastname: Lai
authors.affiliation: Guangdong Key Laboratory of Reproductive Medicine, Guangzhou, Guangdong, 510080, China.
authors.forename: Bing
authors.initials: B
authors.lastname: Cai
authors.affiliation: Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Chen
authors.affiliation: Center for Reproductive Medicine, Key Laboratory for Reproductive Medicine of Guangdong Province, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510140, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Chen
authors.affiliation: Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, China.
authors.forename: Zhichen
authors.initials: Z
authors.lastname: Zhai
authors.affiliation: Program of Stem Cells and Regenerative Medicine, Affiliated Guangzhou Women and Children's Hospital, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510623, China.
authors.forename: Lihua
authors.initials: L
authors.lastname: Huang
authors.affiliation: Department of Ultrasound, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510632, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Li
authors.affiliation: Department of General Surgery, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, China.
authors.forename: Anbin
authors.initials: A
authors.lastname: Hu
authors.affiliation: State Key Laboratory of Ophthalmology, Zhong Shan Ophthalmic Center, Sun Yat-sen University, Guangzhou, 510060, China.
authors.forename: Frank Fuxiang
authors.initials: FF
authors.lastname: Mao
authors.affiliation: Program of Stem Cells and Regenerative Medicine, Affiliated Guangzhou Women and Children's Hospital, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510623, China.|Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, China.|Guangdong Key Laboratory of Reproductive Medicine, Guangzhou, Guangdong, 510080, China.|Department of Biochemistry, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, China.
authors.forename: Andy Peng
authors.initials: AP
authors.lastname: Xiang
authors.affiliation: Department of Pharmacology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, China. taol@mail.sysu.edu.cn.
authors.forename: Liang
authors.initials: L
authors.lastname: Tao
authors.affiliation: Program of Stem Cells and Regenerative Medicine, Affiliated Guangzhou Women and Children's Hospital, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510623, China. liweiq6@mail.sysu.edu.cn.|Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-Sen University, Guangzhou, 510080, China. liweiq6@mail.sysu.edu.cn.|Department of Biochemistry, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080, China. liweiq6@mail.sysu.edu.cn.
authors.forename: Weiqiang
authors.initials: W
authors.lastname: Li
mesh_terms: D049109:Cell Proliferation; D054338:Cell Transdifferentiation / Q000235:genetics*; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D017630:Connexins / Q000235:genetics*; D000066630:Ectopic Gene Expression; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D017629:Gap Junctions / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D000090062:Kruppel-Like Factor 4; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D017630:Connexins; C000715467:KLF4 protein, human; D000090062:Kruppel-Like Factor 4; D014157:Transcription Factors; C082920:connexin 45