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V. Zubaty, M. Valtink, C. Lange, J. Zurdel, V. Doubilei, A.R. Zander, K. Engelmann; Transplantation of Mesenchymal Stem Cells Into RCS Rats for Retinal Repair . Invest. Ophthalmol. Vis. Sci. 2005;46(13):4160.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: The Royal College of Surgeons (RCS) rat, an animal model for inherited retinal dystrophy, undergoes a progressive retinal degeneration during the early postnatal period. Multipotential mesenchymal stem cells (MSC) are assumed to be capable of self–renewal and differentiation into lineages of all three germ layers. Aim of the study was to examine MSC localisation and survival after intraocular and systemic transplantation in normal and dystrophic RCS rats. Methods: Mesenchymal stem cells were isolated from tibiae and femurs of wildtype RCS rats and cultured in DMEM/F12 + 20% preselected FCS. Expanded cells were retrovirally transduced with an eGFP–containing vector and cloned. eGFP–labelled mesenchymal stem cells were injected into the tail vein, vitreous or subretinal space of wildtype and mutant young (2–3 mon) or adult (1 yr) RCS rats. At 1–4 weeks post–transplantation, eyes, lung, heart, liver, spleen and reproductive organs were dissected, fixed in formalin and embedded in paraffin. Tissue sections of 4µm were deparaffinized, treated with proteinase K, and incubated with normal goat serum followed by incubation with a monoclonal chicken antibody directed specifically against GFP. TRITC–labelled rabbit–anti chicken–antibody was used for visualisation, and sections were counterstained with DAPI. In negative controls, the primary antibody was replaced by normal mouse serum. Results: Expression of GFP was evaluated in different tissues at 1–4 weeks post–transplantation. After subretinal or intravitreal injection there was widespread migration of GFP+ cells throughout all retinal layers in normal and mutant rats, with a higher accumulation rate in the degenerated retina of mutant RCS rats. After systemic transplantation, MSC were found in lung, heart and liver. Staining against GFP demarcated retinal autofluorescence from fluorescence signals generated by labelled MSC, confirming the presence of transplanted MSC. Conclusions: Mesenchymal stem cells are capable of integrating into and surviving in the normal and dystrophic retina after local application, and are also capable of migrating and integrating into other organs after systemic injection. The results indicate that MSC have the ability to incorporate into the retina after local application as well as into other organs after systemic injection. Local retinal injection of MSC might thus be a helpful tool in repairing dystrophic lesions in the retina.
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