title: Human limbal biopsy-derived stromal stem cells prevent corneal scarring.
abstract: Conventional allograft therapy for corneal scarring is widespread and successful, but donor tissue is not universally available, and some grafts fail owing to rejection and complications such as endothelial failure. We investigated direct treatment of corneal scarring using autologous stem cells, a therapy that, if successful, could reduce the need for corneal grafts. Mesenchymal cells were expanded from small superficial, clinically replicable limbal biopsies of human cadaveric corneo-scleral rims. Limbal biopsy-derived stromal cells (LBSCs) expanded rapidly in media containing human serum, were highly clonogenic, and could generate spheres expressing stem cell genes (ABCG2, Nestin, NGFR, Oct4, PAX6, and Sox2). Human LBSCs differentiated into keratocytes expressing characteristic marker genes (ALDH3A1, AQP1, KERA, and PTGDS) and organized a thick lamellar stroma-like tissue containing aligned collagen and keratan sulfate proteoglycans when cultured on aligned nanofiber substrata. When engrafted into mouse corneal wounds, LBSCs prevented formation of light-scattering scar tissue containing fibrotic matrix components. The presence of LBSCs induced regeneration of ablated stroma with tissue exhibiting lamellar structure and collagen organization indistinguishable from that of native tissue. Because the limbus can be easily biopsied from either eye of an affected individual and LBSCs capable of corneal stromal remodeling can be expanded under xeno-free autologous conditions, these cells present a potential for autologous stem cell-based treatment of corneal stromal blindness.
authors.affiliation: Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA. L V Prasad Eye Institute, Hyderabad 500034, India.
authors.forename: Sayan
authors.initials: S
authors.lastname: Basu
authors.affiliation: Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Hertsenberg
authors.affiliation: Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA.
authors.forename: Martha L
authors.initials: ML
authors.lastname: Funderburgh
authors.affiliation: Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA.
authors.forename: Michael K
authors.initials: MK
authors.lastname: Burrow
authors.affiliation: Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA.
authors.forename: Mary M
authors.initials: MM
authors.lastname: Mann
authors.affiliation: Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA.
authors.forename: Yiqin
authors.initials: Y
authors.lastname: Du
authors.affiliation: Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA.
authors.forename: Kira L
authors.initials: KL
authors.lastname: Lathrop
authors.affiliation: Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA.
authors.forename: Fatima N
authors.initials: FN
authors.lastname: Syed-Picard
authors.affiliation: Department of Molecular Pharmacology and Physiology, Morsani College of Medicine, University of South Florida, Tampa, FL 33612, USA.
authors.forename: Sheila M
authors.initials: SM
authors.lastname: Adams
authors.affiliation: Department of Molecular Pharmacology and Physiology, Morsani College of Medicine, University of South Florida, Tampa, FL 33612, USA.
authors.forename: David E
authors.initials: DE
authors.lastname: Birk
authors.affiliation: Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA. jlfunder@pitt.edu.
authors.forename: James L
authors.initials: JL
authors.lastname: Funderburgh
mesh_terms: D000328:Adult; D000818:Animals; D001706:Biopsy; D002454:Cell Differentiation; D002921:Cicatrix / Q000517:prevention & control; D003094:Collagen / Q000737:chemistry; D003315:Cornea / Q000378:metabolism / Q000473:pathology*; D016039:Corneal Transplantation / Q000379:methods*; D003470:Culture Media / Q000737:chemistry; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D005786:Gene Expression Regulation; D006801:Humans; D007632:Keratan Sulfate / Q000737:chemistry; D015603:Keratinocytes / Q000166:cytology; D051379:Mice; D008810:Mice, Inbred C57BL; D008875:Middle Aged; D011509:Proteoglycans / Q000737:chemistry; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology*; D017154:Stromal Cells / Q000166:cytology*
chemical_list: D003470:Culture Media; D011509:Proteoglycans; D003094:Collagen; D007632:Keratan Sulfate