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Yuan Dong, Kaushali Thakore-Shah, Elfren Ray Baclagon, Sophie Xiaohui Deng; Isolation and Expansion of Porcine Non-pigmented Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2016;57(12):217.
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To develop a method for the isolation, and the expansion of non-pigmented epithelium (NPE).
Intact rings of NPE and pigmented epithelium (PE, NPE+PE) or NPE alone were isolated from adult porcine eyes. The structure of NPE+ PE or NPE tissue was examined by histochemistry. Single cell cultures of NPE were maintained for up to 10 passages in either DMEM F12 or SFM base media supplemented with growth factors and fetal bovine serum. Immunohistochemistry was performed on cultured NPE cells as well as frozen tissue sections to examine the expression of several NPE markers.
All media conditions tested supported NPE growth for more than 10 several passages. Cells cultured in SFM base medium with 5% FBS appeared to be more homogeneous and maintained as a single monolayer with typical epithelial cobblestone appearance. Robust expression and membrane localization of the tight junction protein ZO-1 and pump protein Na-K ATPase were detected in NPE cells. Expression of NPE markers were maintained as well.
We have developed a reproducible method to isolate and culture NPE cells without losing their phenotype. The NPE cells cultured under this condition would be valuable for future studies.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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