Gingival Fibroblasts as Autologous Feeders for Induced Pluripotent Stem Cells

Human gingival fibroblasts (hgfs) present an attractive source of induced pluripotent stem cells (ipscs),which are expected to be a powerful tool for regenerative dentistry. however,problems to be addressed prior to clinical application include the use of animal-derived feeder cells for cultures. the aim of this study was to establish an autologous hgf-derived ipsc (hgf-ipsc) culture system by evaluating the feeder ability of hgfs. in both serum-containing and serum-free media,hgfs showed higher proliferation than human dermal fibroblasts (hdfs). three hgf strains were isolated under serum-free conditions,although 2 showed impaired proliferation. when hgf-ipscs were transferred onto mitomycin c-inactivated hgfs,hdfs,or mouse-derived snl feeders,hgf and snl feeders were clearly hgf-ipsc supportive for more than 50 passages,whereas hdf feeders were only able to maintain undifferentiated hgf-ipsc growth for a few passages. after 20 passages on hgf feeders,embryonic stem cell marker expression and cpg methylation at the nanog and oct3/4 promoters were similar for hgf-ipscs cultured on hgf and snl feeder cells. long-term cultures of hgf-ipscs on hgf feeders sustained their normal karyotype and pluripotency. on hgf feeders,hgf-ipsc colonies were surrounded by many colony-derived fibroblast-like cells,and the size of intact colonies at 7 d after passage was significantly larger than that on snl feeders. allogeneic hgf strains also maintained hgf-ipscs for 10 passages. compared with hdfs,hgfs showed a higher production of laminin-332,laminin α5 chain,and insulin-like growth factor-ii,which have been reported to sustain the long-term self-renewal of pluripotent stem cells. these results suggest that hgfs possess an excellent feeder capability and thus can be used as alternatives to conventional mouse-derived snl and hdf feeders. in addition,our findings suggest that hgf feeders are promising candidates for animal component-free ex vivo expansion of autologous hgf-ipscs,thus providing an important step toward the future therapeutic application of hgf-ipscs. © international & american associations for dental research.