Methods for culturing retinal pigment epithelial cells: a review of current protocols and future recommendations

Abstract

The retinal pigment epithelium is an important part of the vertebrate eye, particularly in studying the causes and possible treatment of age-related macular degeneration. The retinal pigment epithelium is difficult to access in vivo due to its location at the back of the eye, making experimentation with age-related macular degeneration treatments problematic. An alternative to in vivo experimentation is cultivating the retinal pigment epithelium in vitro, a practice that has been going on since the 1970s, providing a wide range of retinal pigment epithelial culture protocols, each producing cells and tissue of varying degrees of similarity to natural retinal pigment epithelium. The purpose of this review is to provide researchers with a ready list of retinal pigment epithelial protocols, their effects on cultured tissue, and their specific possible applications. Protocols using human and animal retinal pigment epithelium cells, derived from tissue or cell lines, are discussed, and recommendations for future researchers included.

Human retinal pigment epithelial cells prefer proline as a nutrient and transport metabolic intermediates to the retinal side

“…RPE culture is an excellent in vitro model to study RPE function and AMD (17,21). In this study, we used targeted metabolomics to study how the RPE consumes nutrients.…”

Development and characterization of an in vitro system of the human retina using cultured cell lines

“…Often disputed in literature, is the use of cell lines in model design due to the presence of developmental abnormalities . The prolonged culture of both ARPE‐19 and MIO‐M1 can result in abnormalities in characteristic features of the cell such as the loss of pigmentation in ARPE‐19 and MIO‐M1 exhibiting progenitor characteristics .…”

“…The prolonged culture of both ARPE‐19 and MIO‐M1 can result in abnormalities in characteristic features of the cell such as the loss of pigmentation in ARPE‐19 and MIO‐M1 exhibiting progenitor characteristics . However, within our study design we have allowed for maturation and low passage numbers to ensure manipulation of culture conditions will result in minimal difference from parent tissue . The superior use of an alternative cell source that is, primary cell sources will yield a truer representation of the native tissue.…”

“…Often disputed in literature, is the use of cell lines in model design due to the presence of developmental abnormalities. 17,26 The prolonged culture of both ARPE-19 and MIO-M1 can result in abnormalities in characteristic features of the cell such as the loss of pigmentation in ARPE-19 and MIO-M1 exhibiting progenitor characteristics. 27 However, within our study design we have allowed for maturation and low passage numbers to ensure manipulation of culture conditions will result in minimal difference from parent tissue.…”

“…27 However, within our study design we have allowed for maturation and low passage numbers to ensure manipulation of culture conditions will result in minimal difference from parent tissue. 26 The superior use of an alternative cell source that is, primary cell sources will yield a truer representation of the native tissue. However, it can also produce poor efficiency of the model in practice allowing for restricted use of repeats and minimize use in pharmacological testing.…”