Guest Editors
Aparna Lakkaraju
University of California, San Francisco
Dr. Aparna Lakkaraju is a Professor of Ophthalmology at the University of California, San Francisco. Research in the Lakkaraju laboratory focuses on mechanisms that maintain homeostasis in the retina and retinal pigment epithelium (RPE) with the goal of developing effective therapies for inherited and age-related macular degeneration (AMD). The RPE, which performs numerous functions indispensable for vision, is a key site of injury in macular degeneration. Using multidisciplinary approaches such as advanced live cells, the Lakkaraju lab has identified novel therapeutics that preserve RPE health and function.
Li Xuan Tan
University of California, San Francisco
Dr. Li Xuan Tan is a postdoctoral scholar in the Lakkaraju Lab with 10 years of experience in the field of RPE cell biology. She obtained her PhD in Pharmaceutical Sciences at the University of Wisconsin-Madison in 2018. She worked on regulating the complement system in the retinal pigment epithelium (RPE) and therapeutic strategy for restoring protective mechanisms against complement-mediated injury. Her current research focuses on regulating phagocytosis and clearance of photoreceptor outer segments in the RPE.
Collection Overview
The retinal pigment epithelium (RPE) is a monolayer of polarized postmitotic cells between the light-sensing photoreceptors and the choriocapillaris. The RPE performs numerous indispensable functions in maintaining photoreceptor health and vision. Damage to the RPE is an initiating factor for blinding macular degeneration. The ability of the RPE to perform its functions is tightly linked to its state of differentiation and polarity. Mature, polarized RPE have highly specialized apical and basolateral domains demarcated by tight junctions. The RPE also has distinct subcellular distributions of organelles, such as apically localized melanosomes and distinct apical and basal endosomes.
The polarity of the RPE facilitates the directed release of growth factors and extracellular matrix components to the apical or basal side of the RPE. Tight junctions between the RPE form the outer blood-retina barrier that regulates the transport of ions, nutrients, and metabolites into and out of the retina. To study the RPE in vitro, primary cultures that recapitulate the polarity and differentiation characteristics of the RPE in situ are essential. This approach requires standardized protocols for generating RPE cultures from laboratory animals and human donors. The lack of standardized culture conditions makes it difficult to obtain RPE cultures that resemble native RPE and could result in low reproducibility between studies.
This Methods Collection will include protocols for obtaining primary RPE cultures from mouse, rat, porcine, and human donors. Although stem cell-derived RPEs are used to model RPE diseases, they face variability issues and varying extents of epigenetic reprogramming, which could limit their widespread use. In this regard, primary adult RPE cultures established from lab animals and human donors would better recapitulate age-related retinal disease. This means that this collection will focus on protocols for establishing primary RPE cultures from experts in the field.
