title: Tissue engineering of corneal stromal layer with dermal fibroblasts: phenotypic and functional switch of differentiated cells in cornea.
abstract: Previously, we successfully engineered a corneal stromal layer using corneal stromal cells. However, the limited source and proliferation potential of corneal stromal cells has driven us to search for alternative cell sources for corneal stroma engineering. Based on the idea that the tissue-specific environment may alter cell fate, we proposed that dermal fibroblasts could switch their phenotype to that of corneal stromal cells in the corneal environment. Thus, dermal fibroblasts were harvested from newborn rabbits, seeded on biodegradable polyglycolic acid (PGA) scaffolds, cultured in vitro for 1 week, and then implanted into adult rabbit corneas. After 8 weeks of implantation, nearly transparent corneal stroma was formed, with a histological structure similar to that of its native counterpart. The existence of cells that had been retrovirally labeled with green fluorescence protein (GFP) demonstrated the survival of implanted cells. In addition, all GFP-positive cells that survived expressed keratocan, a specific marker for corneal stromal cells, and formed fine collagen fibrils with a highly organized pattern similar to that of native stroma. However, neither dermal fibroblast-PGA construct pre-incubated in vitro for 3 weeks nor chondrocyte-PGA construct could form transparent stroma. The results demonstrated that neonatal dermal fibroblasts could switch their phenotype in the new tissue environment under restricted conditions. The functional restoration of corneal transparency using dermal fibroblasts suggests that they could be an alternative cell source for corneal stroma engineering.
authors.affiliation: Shanghai Jiao Tong University School of Medicine, Plastic and Reconstructive Surgery, Shanghai 9th People's Hospital, Shanghai, China.
authors.forename: Yan Qing
authors.initials: YQ
authors.lastname: Zhang
authors.forename: Wen Jie
authors.initials: WJ
authors.lastname: Zhang
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.forename: Xiao Jie
authors.initials: XJ
authors.lastname: Hu
authors.forename: Guang Dong
authors.initials: GD
authors.lastname: Zhou
authors.forename: Lei
authors.initials: L
authors.lastname: Cui
authors.forename: Yilin
authors.initials: Y
authors.lastname: Cao
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002454:Cell Differentiation; D002470:Cell Survival; D003315:Cornea / Q000166:cytology* / Q000378:metabolism / Q000601:surgery; D020405:Dermis / Q000166:cytology / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D011100:Polyglycolic Acid / Q000737:chemistry; D011817:Rabbits; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry
chemical_list: D049452:Green Fluorescent Proteins; D011100:Polyglycolic Acid