Abstract
Dry eye is a complicated ocular surface disease whose exact pathogenesis is not yet fully understood. For the therapeutic evaluation and pathogenesis study of dry eye, we established an in vitro three-dimensional (3D) coculture model for the ocular surface. It is composed of rabbit conjunctival epithelium and lacrimal gland cell spheroids, and recapitulates the aqueous and mucin layers of the tear film. We first investigated the culture conditions for both cell types to optimize their secretory functions, by employing goblet cell enrichment, air-lifting culture, and 3D spheroid formation techniques. The coculture of the two cell components leads to elevated secretion and higher expression of tear secretory markers. We also compared several coculture systems, and found that direct cell contact between the two cell types significantly increased tear secretion. Inflammation was induced to mimic dry eye disease in the coculture model system. Our results showed that the coculture system provides a more physiologically relevant therapeutic response compared to monocultures. Our work provides a complex 3D model as a recapitulation of the ocular surface and tear film system, which can be further developed as a model for dry eye disease and therapeutic evaluation.
Highlights
The ocular surface is the outermost layer of the eye
Enriched conjunctival epithelial cells (CECs) expressed more epithelial markers in culture and secreted more mucosubstance when compared to unseparated control, as confirmed by cytokeratin 4 (CK4), Periodic acid-Schiff (PAS), and mucin 5AC (MUC5AC) stainings, respectively (Fig. 2B)
The mucin-aqueous layer in the tear film is of essential importance in protecting the ocular surface epithelium, and is frequently damaged in dry eye disease
Summary
The ocular surface is the outermost layer of the eye. Together with the tear film, it protects the refractive surface and enables sharp vision[1]. Considering the ocular surface and tear film, researchers have been investigating the in vitro modeling of several components individually, including conjunctival epithelium[17] and lacrimal glands[18, 19] In these models, cell morphology and phenotype resembled the equivalent cells in vivo. Combining these cells types to create a functional tear film has not been studied To address this gap, we developed a complex three-dimensional (3D) in vitro model for the ocular surface. We developed a complex three-dimensional (3D) in vitro model for the ocular surface This model contains primary rabbit conjunctival epithelial cells (CECs) and lacrimal gland (LG) cell spheroids, to recapitulate the aqueous and mucin layers of the tear film. We engineered a model system for both a healthy and diseased ocular surface amenable to studying disease pathogenesis and therapeutic screening
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