Reconstruction of damaged corneal epithelium using Venus-labeled limbal epithelial stem cells and tracking of surviving donor cells.
Limbal epithelial stem cells are responsible for the self-renewal and replenishment of the corneal epithelium. Although it is possible to repair the ocular surface using limbal stem cell transplantation, the mechanisms behind this therapy are unclear. To investigate the distribution of surviving donor cells in a reconstructed corneal epithelium, we screened a Venus-labeled limbal stem cell strain in goats. Cells were cultivated on denuded human amniotic membrane for 21 days to produce Venus-labeled corneal epithelial sheets. The Venus-labeled corneal epithelial sheets were transplanted to goat models of limbal stem cell deficiency. At 3 months post-surgery, the damaged corneal epithelia were obviously improved in the transplanted group compared with the non-transplanted control, with the donor cells still residing in the reconstructed ocular surface epithelium. Using Venus as a marker, our results indicated that the location and survival of donor cells varied, depending on the corneal epithelial region. Additionally, immunofluorescent staining of the reconstructed corneal epithelium demonstrated that many P63(+) cells were unevenly distributed among basal and suprabasal epithelial layers. Our study provides a new model, and reveals some of the mechanisms involved in corneal epithelial cell regeneration research.
College of Veterinary Medicine, Northwest A&F University, Shaanxi Stem Cell Engineering and Technology Center, Yangling 712100, China.
Ji-Qing
JQ
Yin
Wen-Qiang
WQ
Liu
Chao
C
Liu
Yi-Hua
YH
Zhang
Jin-Lian
JL
Hua
Wei-Shuai
WS
Liu
Zhong-Ying
ZY
Dou
An-Min
AM
Lei
D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D000650:Amnion / Q000166:cytology; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D015415:Biomarkers / Q000378:metabolism; D002470:Cell Survival; D002478:Cells, Cultured; D003316:Corneal Diseases / Q000601:surgery*; D065306:Corneal Injuries; D019573:Epithelium, Corneal / Q000473:pathology* / Q000601:surgery; D005131:Eye Injuries / Q000601:surgery*; D005456:Fluorescent Dyes; D005822:Genetic Vectors; D006041:Goats; D039641:Integrin beta Chains / Q000378:metabolism; D053539:Keratin-19 / Q000378:metabolism; D016850:Limbus Corneae / Q000166:cytology*; D008164:Luminescent Proteins / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D013194:Staining and Labeling; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism; D014019:Tissue Donors; D014162:Transfection; D014184:Transplantation, Homologous; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D015415:Biomarkers; D005456:Fluorescent Dyes; D039641:Integrin beta Chains; D053539:Keratin-19; D008164:Luminescent Proteins; D025521:Tumor Suppressor Proteins; C054562:yellow fluorescent protein, Bacteria
LSCT; Venus-labeled corneal epithelium sheets; limbal epithelial stem cells; reconstructed corneal epithelium