title: Substrate-mediated reprogramming of human fibroblasts into neural crest stem-like cells and their applications in neural repair.
abstract: Cell- and gene-based therapies have emerged as promising strategies for treating neurological diseases. The sources of neural stem cells are limited while the induced pluripotent stem (iPS) cells have risk of tumor formation. Here, we proposed the generation of self-renewable, multipotent, and neural lineage-related neural crest stem-like cells by chitosan substrate-mediated gene transfer of a single factor forkhead box D3 (FOXD3) for the use in neural repair. A simple, non-toxic, substrate-mediated method was applied to deliver the naked FOXD3 plasmid into human fibroblasts. The transfection of FOXD3 increased cell proliferation and up-regulated the neural crest marker genes (FOXD3, SOX2, and CD271), stemness marker genes (OCT4, NANOG, and SOX2), and neural lineage-related genes (Nestin, β-tubulin and GFAP). The expression levels of stemness marker genes and neural crest maker genes in the FOXD3-transfected fibroblasts were maintained until the fifth passage. The FOXD3 reprogrammed fibroblasts based on the new method significantly rescued the neural function of the impaired zebrafish. The chitosan substrate-mediated delivery of naked plasmid showed feasibility in reprogramming somatic cells. Particularly, the FOXD3 reprogrammed fibroblasts hold promise as an easily accessible cellular source with neural crest stem-like behavior for treating neural diseases in the future.
authors.affiliation: Institute of Polymer Science and Engineering, National Taiwan University, Taipei, Taiwan.
authors.forename: Ting-Chen
authors.initials: TC
authors.lastname: Tseng
authors.affiliation: Institute of Polymer Science and Engineering, National Taiwan University, Taipei, Taiwan.
authors.forename: Fu-Yu
authors.initials: FY
authors.lastname: Hsieh
authors.affiliation: Division of Plastic and Reconstructive Surgery, Department of Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.
authors.forename: Niann-Tzyy
authors.initials: NT
authors.lastname: Dai
authors.affiliation: Institute of Polymer Science and Engineering, National Taiwan University, Taipei, Taiwan; Center of Tissue Engineering and 3D Printing, National Taiwan University, Taipei, Taiwan. Electronic address: shhsu@ntu.edu.tw.
authors.forename: Shan-Hui
authors.initials: SH
authors.lastname: Hsu
mesh_terms: D000328:Adult; D000818:Animals; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D000067470:Cellular Reprogramming Techniques / Q000379:methods*; D048271:Chitosan / Q000737:chemistry; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D051858:Forkhead Transcription Factors / Q000235:genetics*; D006801:Humans; D008297:Male; D009432:Neural Crest / Q000166:cytology* / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D055495:Neurogenesis; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D014162:Transfection / Q000379:methods*; D015027:Zebrafish
chemical_list: C503514:FOXD3 protein, human; D051858:Forkhead Transcription Factors; D048271:Chitosan
keywords: Forkhead box D3 (FOXD3); Neural crest; Neural repair; Substrate-mediated cell reprogramming