INTRODUCTION: Multilamellar bodies (MLBs) are concentric cytoplasmic membranes which form through an autophagy-dependent mechanism. In the cornea, the presence of MLBs is associated with Schnyder corneal dystrophy (SCD). Ex vivo 3D modelling of the corneal stroma and SCD can help study pathogenesis and resolution of the disorder. METHODS: Corneal stroma explants were isolated from cadavers and cultivated long-term for more than 3 months to achieve spontaneous 3D outgrowth of corneal stroma-derived mesenchymal stem-like cells (CSMSCs). The 3D tissues were then examined by transmission electron microscopy (TEM) for presence of MLBs, and by immunofluorescent labelling against markers for autophagy (p62, LC3). Autophagy was induced by classical serum starvation or rapamycin (RAP) treatment (50 nM), and inhibited by the autophagy inhibitor 3-methyladenine (3-MA, 10 mM) for 24 hours. RESULTS: CSMSCs can form spontaneously 3D outgrowths over a 3-4 weeks period, depositing their own extracellular matrix containing collagen I. TEM confirmed the presence of MLBs in the long-term (>3 months) 3D cultures, which became more abundant under starvation and RAP treatment, and decreased in number under autophagy inhibition with 3-MA. The presence of autophagy and its disappearance could be confirmed by an inversely related increase and decrease in the expression of LC3 and p62, respectively. CONCLUSIONS: MLB formation in long-standing CSMSC cultures could serve as a potential ex vivo model for studying corneal stroma diseases, including SCD. Inhibition of autophagy can decrease the formation of MLBs, which may lead to a novel treatment of the disease in the future.

• MeSH
• adenin analogy a deriváty   farmakologie   MeSH
• anatomické modely MeSH
• autofagie * účinky léků   MeSH
• buněčná inkluze patologie   ultrastruktura   MeSH
• dědičné dystrofie rohovky patologie   patofyziologie   MeSH
• dospělí MeSH
• extracelulární matrix metabolismus   ultrastruktura   MeSH
• fluorescenční protilátková technika MeSH
• kultivované buňky MeSH
• lidé středního věku MeSH
• lidé MeSH
• mezenchymální kmenové buňky MeSH
• mrtvola MeSH
• rohovka metabolismus   MeSH
• senioři MeSH
• stroma rohovky patologie   patofyziologie   ultrastruktura   MeSH
• transmisní elektronová mikroskopie MeSH
• zobrazování trojrozměrné MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH