Abstract
Background: The complement system (CS) plays a role in the pathogenesis of a number of ocular diseases, including diabetic retinopathy (DR), glaucoma, uveitis, and age-related macular degeneration (AMD). Given that many of the complex eye-related degenerative diseases have limited treatment opportunities, we aimed to mimic the in vivo retinal degenerative process by developing a relevant co-culture system.Method and materials: The adult porcine retina was co-cultured with the spontaneously arising human retinal pigment epithelial cells-19 (ARPE-19).Results: Inflammatory activity was found after culture and included migrating microglial cells, gliosis, cell death, and CS activation (demonstrated by a minor increase in the secreted anaphylotoxin C3a in co-culture). CS components, including C1q, C3, C4, soluble C5b-9, and the C5a receptor, were expressed in the retina and/or ARPE cells after culture. C1q, C3, and CS regulators such as C4 binding protein (C4BP), factor H (CFH), and factor I (CFI) were secreted after culture.Discussion: Thus, our research indicates that this co-culturing system may be useful for investigations of the CS and its involvement in experimental neurodegenerative diseases.
Highlights
The complement system (CS), an essential element of innate immunity, is an organized complex network of more than 50 soluble and surface proteins, primarily produced in the liver, that are activated through proteolytic cascades [1,2]
Retinal pigment epithelium 65 (RPE65) was expressed in the post-confluent arising human retinal pigment epithelial (ARPE)-cells (Figure 2)
The experimental co-culturing system used in our experiments, i.e. culturing adult porcine retinas with post-confluent ARPE cells, offers a controlled in vitro system that can mimic a diverse set of retinal diseases
Summary
The complement system (CS), an essential element of innate immunity, is an organized complex network of more than 50 soluble and surface proteins, primarily produced in the liver, that are activated through proteolytic cascades [1,2]. The AP is in a continuous active low-level state under physiological conditions and needs to be tightly controlled by endogenous regulators to avoid tissue damage [4]. Both soluble and membrane-bound regulators are needed to control the CS at various levels. CS components, including C1q, C3, C4, soluble C5b-9, and the C5a receptor, were expressed in the retina and/or ARPE cells after culture. Discussion: our research indicates that this co-culturing system may be useful for investigations of the CS and its involvement in experimental neurodegenerative diseases
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