Comparison of retinal pigment epithelium cell preparations from the bovine eye.
Retinal pigment epithelium (RPE) cells were collected from bovine eyes using a new method. The cells were harvested by vortexing the RPE and underlying choroid in 0.05 M citrate phosphate buffer, pH 5. RPE cells recovered by this method were compared to a standard method by microscopic examination of cell integrity, estimation of total protein, and assay of 11-cis and all-trans retinyl ester hydrolase (REH) activities. Results suggest that this method collects RPE cells of good integrity and with a significantly higher protein yield than the conventional method. Additionally, a much higher retinyl ester hydrolase activity was noted. Therefore we propose that this procedure offers a new and convenient method in the collection of RPE proteins for certain purposes such as enzyme purification.
Sanders-Sanchez
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Division of Life Sciences, University of Texas, San Antonio 78285.
Malsbury
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Tsin
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D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism; D002417:Cattle; D002452:Cell Count; D002469:Cell Separation / Q000379:methods; D005069:Evaluation Studies as Topic; D005136:Eye Proteins / Q000378:metabolism; D010857:Pigment Epithelium of Eye / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism; D013237:Stereoisomerism
D005136:Eye Proteins; D002265:Carboxylic Ester Hydrolases; C022523:retinyl esterase