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M. Omoto, H. Miyashita, K. Higa, K. Tsubota, S. Shimmura; The Use of Human Mesenchymal Stem Cells as Feeder Cells in Cultivated Corneal Epithelial Cell Sheets. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5748.
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© ARVO (1962-2015); The Authors (2016-present)
Corneal epithelial cell sheets, widely used to reconstruct the ocular surface, are mostly co-cultured with 3T3 feeder cells established from mouse embryo. To evade the risk of contagion associated with xenografting, we used human mesenchymal stem cells as feeder cells, and evaluated the quality of the epithelial cell sheets.
Human bone marrow mesenchymal stem cells (Marrow Adherent Stem Cells: MASC) were cultured and treated with mitomycin C to use as feeder cells. Human limbal epithelial cells were obtained from US eye bank corneas and co-cultured at a low density with MASC feeder cells or 3T3 feeder cells to compare colony forming efficiency. Limbal epithelial cells were cultured on MASC or 3T3 feeder cells at liquid-air interface to allow stratification, and stratified epithelial sheets were analyzed by immunohistochemistry. Rabbit limbal epithelial cells were cultivated with MASC feeder cells, and stratified epithelial cell sheets were transplanted to the ocular surface of limbal deficient rabbits. Epithelial grafts were observed by slit lamp microscopy for 4 weeks after surgery, and then harvested for histology and immunohistochemistry.
The colony-forming efficiency of human limbal epithelial cells was similar in both groups. Stratified cell sheets were successfully cultivated with MASC feeder cells, expressing K3 and K15. Epithelial grafts maintained a clear ocular surface in rabbits for 4 weeks, and exhibited the corneal epithelium phenotype.
MASC feeder cells can be used as a substitution for 3T3 in the cultivation of transplantable epithelial sheets.
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