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Benedicte Tougeron Brousseau, Julie Gueudry, Marek Lamacz, Jean-Pierre Vannier, Marc Muraine; Does Rabbit Bone Marrow -derived Cells Implantation Contribute To Corneal Epithelial Repair?. Invest. Ophthalmol. Vis. Sci. 2011;52(14):2030. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
Bone marrow-derived stem cells were reported to differentiate in epithelial cells in vitro and in vivo. Amniotic membrane is a good basement membrane for cultivating limbal corneal epithelial cells. We explore the potential of adherent bone marrow-derived stem cells to provide a therapy for limbal stem cells insufficiency after transfer on human amniotic membrane and we analyze their outcome in vivo.
Rabbit adherent bone marrow stem cells (mesenchymal stem cell, MSC) were expanded and transferred on denuded amniotic membrane (MA). To trace stem cells, we infected them with a retroviral vector encoding the Green Fluorescent Protein (GFP). Optimal conditions for expansion of rabbit adherent bone marrow cells were determined. A confluent culture of MSC was established on MA. Bone marrow cells on MA were analysed by HES coloration and epithelial differentiation were analysed by immunohistochemistry. MA were autologous transplanted onto rabbit eyes injured by a lamellar keratectomy extending 3 mm outside the limbus, performed one month ago. After one month, animals were sacrificed. Dissected corneas were analysing with HES coloration and fluorescent microscopy.
After transplantion on the rabbit eyes with limbal stem cells insufficiency, GFP fluorescence was detect in the cornea. The examination with fluorescent microscopy has showed a presence of MSC labelled with GFP in the cornea stroma. Nevertheless, their was no amelioration of the ocular surface disorder in microscopic analysis. Moreover, majority of MSC were lysed after 3 weeks of the transplantation.
Autologous transplantion of MSC is feasible by using amniotic membrane as a basement membrane. The transplantation MSC does not seem to ameliorate the ocular surface disorder in our model probably due to their lysis. It would be interesting to research the reasons of this lyse and evaluate the possibilities of gene transfer in MSC for therapeutic strategies.
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