Methods in Enzymology Vol.190 Retinoids, Part B: Cell Differentiation and Clinical Applications

Retinoid metabolism in the retinal pigment epithelium (RPE), with respect to the visual cycle, encompasses several steps: uptake of retinol, intracellular transport, acylation, isomerization, oxidation, and secretion of retinoid. To study such a complex set of metabolic pathways in RPE cells, the development of a reliable in vitro system would be highly desirable. Investigations on the multifaceted retinoid metabolism in RPE cells in vitro require isolated RPE cells in which the complexity of cellular organization is preserved to a high extent and which are viable and metabolically active. To meet these requirements of physiological fitness, the isolation of bovine RPE cells and the in vitro incubation conditions have been optimized by applying a variety of criteria. These criteria included morphology (ultrastructure of the cells), viability (exclusion of viability stains and retention of small cellular proteins), and metabolic activity (energy charge). This chapter describes an approach to study retinoid metabolism in isolated bovine RPE cells during short-term in vitro incubation.

• Series: Methods in Enzymology (Book 190)
• Hardcover: 488 pages
• Publisher: Academic Press;
  
• Language: English
• ISBN-10: 0121820912
• ISBN-13: 978-0121820916
• Product Dimensions: 1.2 x 6.5 x 9.8 inches

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