Substrate-mediated reprogramming of human fibroblasts into neural crest stem-like cells and their applications in neural repair
Journal
Biomaterials
Journal Volume
102
Pages
148-161
Date Issued
2016
Author(s)
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. ? 2016 Elsevier Ltd
Subjects
Forkhead box D3 (FOXD3); Neural crest; Neural repair; Substrate-mediated cell reprogramming
Other Subjects
Cell culture; Cell proliferation; Cells; Chitin; Chitosan; Cytology; Disease control; DNA; Fibroblasts; Gene expression; Gene therapy; Gene transfer; Genes; Repair; Stem cells; Cell reprogramming; Expression levels; Forkhead box D3 (FOXD3); Human fibroblast; Neural crests; Neural functions; Neural stem cell; Neurological disease; Molecular biology; beta tubulin; CD271 antigen; chitosan; nestin; octamer transcription factor 4; transcription factor; transcription factor FOXD3; transcription factor NANOG; transcription factor Sox2; unclassified drug; chitosan; forkhead transcription factor; FOXD3 protein, human; adult; Article; cell proliferation; cell renewal; cell viability; controlled study; feasibility study; fibroblast; gene expression; gene transfer; genetic transfection; human; human cell; marker gene; neural crest cell; neural stem cell; neurologic disease; nonhuman; nuclear reprogramming; priority journal; somatic cell; upregulation; zebra fish; administration and dosage; animal; cell culture; cell reprogramming technique; chemistry; cytology; fibroblast; genetics; male; metabolism; nervous system development; neural crest; neural stem cell; nuclear reprogramming; plasmid; procedures; Adult; Animals; Cells, Cultured; Cellular Reprogramming; Cellular Reprogramming Techniques; Chitosan; Fibroblasts; Forkhead Transcription Factors; Humans; Male; Neural Crest; Neural Stem Cells; Neurogenesis; Plasmids; Transfection; Zebrafish
Type
journal article