title: Generation and periodontal differentiation of human gingival fibroblasts-derived integration-free induced pluripotent stem cells.
abstract: Induced pluripotent stem cells (iPSCs) have been recognized as a promising cell source for periodontal tissue regeneration. However, the conventional virus-based reprogramming approach is associated with a high risk of genetic mutation and limits their therapeutic utility. Here, we successfully generated iPSCs from readily accessible human gingival fibroblasts (hGFs) through an integration-free and feeder-free approach via delivery of reprogramming factors of Oct4, Sox2, Klf4, L-myc, Lin28 and TP53 shRNA with episomal plasmid vectors. The iPSCs presented similar morphology and proliferation characteristics as embryonic stem cells (ESCs), and expressed pluripotent markers including Oct4, Tra181, Nanog and SSEA-4. Additionally, these cells maintained a normal karyotype and showed decreased CpG methylation ratio in the promoter regions of Oct4 and Nanog. In vivo teratoma formation assay revealed the development of tissues representative of three germ layers, confirming the acquisition of pluripotency. Furthermore, treatment of the iPSCs in vitro with enamel matrix derivative (EMD) or growth/differentiation factor-5 (GDF-5) significantly up-regulated the expression of periodontal tissue markers associated with bone, periodontal ligament and cementum respectively. Taken together, our data demonstrate that hGFs are a valuable cell source for generating integration-free iPSCs, which could be sequentially induced toward periodontal cells under the treatment of EMD and GDF-5.
authors.affiliation: Department of Periodontology, School and Hospital of Stomatology, Peking University, 22 South Avenue Zhong-Guan-Cun, Beijing 100081, China; Peking University Stem Cell Research Center and Department of Cell Biology, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191, China.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Yin
authors.affiliation: Peking University Stem Cell Research Center and Department of Cell Biology, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Periodontology, School and Hospital of Stomatology, Peking University, 22 South Avenue Zhong-Guan-Cun, Beijing 100081, China.
authors.forename: Jingwen
authors.initials: J
authors.lastname: Li
authors.affiliation: Faculty of Dentistry, The University of Hong Kong, 34 Hospital Road, Hong Kong SAR, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Li
authors.affiliation: Peking University Stem Cell Research Center and Department of Cell Biology, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191, China.
authors.forename: Yinan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Peking University Stem Cell Research Center and Department of Cell Biology, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191, China. Electronic address: jhwen@bjmu.edu.cn.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Wen
authors.affiliation: Department of Periodontology, School and Hospital of Stomatology, Peking University, 22 South Avenue Zhong-Guan-Cun, Beijing 100081, China. Electronic address: kqluanqx@126.com.
authors.forename: Qingxian
authors.initials: Q
authors.lastname: Luan
mesh_terms: D000293:Adolescent; D000328:Adult; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation; D019175:DNA Methylation; D005347:Fibroblasts / Q000166:cytology*; D005881:Gingiva / Q000166:cytology*; D055428:Growth Differentiation Factor 5 / Q000378:metabolism*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D007621:Karyotyping; D000090062:Kruppel-Like Factor 4; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D012038:Regeneration; D013724:Teratoma; D015854:Up-Regulation; D055815:Young Adult
chemical_list: C525893:GDF5 protein, human; D055428:Growth Differentiation Factor 5; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D000469:Alkaline Phosphatase
keywords: Differentiation; Enamel matrix derivative; Growth/differentiation factor-5; Human gingival fibroblasts; Induced pluripotent stem cells; Periodontal regeneration