Background: This infrastructure delivers biological material necessary for several research projects to Vision Health Research Network investigators (VHRN).
Methods: Héma-Québec is the organism in charge obtaining consent and retrieving donor eyes for patient treatment or for research. In Quebec City, donor eyes are sent to the eye bank of the “Centre Universitaire d’Ophtalmologie” (CUO) of Saint-Sacrement hospital. Technicians at the eye bank evaluate the quality of the tissues. Those unfit for graft are transferred to the infrastructure where the coordinator encodes samples prior to their distribution.
Results: Between 2013 and 2017, 27 fundamental investigators, clinical investigators and collaborators supported by 60 students, trainees and laboratory assistants used this infrastructure to move forward their projects. Since 2013, results from those projects generated 21 scientific publications and 232 presentations. The infrastructure helped VHRN investigators obtain near 4 million dollars in grants from many organisms (CIHR, NSERC, Foundations, etc.). These grants allowed recruitment and formation of highly qualified personnel. Last year (April 2016 to March 2017), 189 corneas and 23 eyes transited through the infrastructure.
Conclusions: This infrastructure is available for all investigators that are members of the VHRN. Many original projects have been elaborated thanks to the human ocular tissues provided by this infrastructure. These projects will advance our knowledge in vision health. A better understanding of eye functions will lead to new treatments for eye diseases.
Background: The goal of this study was to engineer an epithelialized and endothelialized pigmented choroidal substitute using the self-assembly approach of tissue engineering.
Methods: Cells from human choroids were isolated and cultured. Culture purity was assessed using immunostaining (CD31, HMB45, vimentin, keratins 8/18). To engineer the choroid, fibroblasts were cultured in the presence of serum and ascorbic acid to promote extracellular matrix (ECM) assembly. Endothelial cells, melanocytes or RPE cells were separately seeded on the stromal substitutes. Choroidal substitutes were further characterized by histology, mass spectrometry, immunostaining, and compared to native human choroids.
Results: The technique used to isolate choroidal cells yielded pure cultures of fibroblasts, melanocytes and vascular endothelial cells. The stromal substitutes engineered using the self-assembly approach were composed of collagen (types I, VI, XII and XIV), proteoglycans (decorin, lumican) and other ECM proteins. Protein expression was confirmed using immunostaining. Endothelial cells spontaneously assembled into capillary-like structures and vascular networks when cocultured with fibroblast-containing ECM sheets.
Conclusions: This study shows that the self-assembly approach of tissue engineering can be used to reconstruct a choroid using native cells. This model represents a unique tool to better understand the crosstalk between the different choroidal cell types and cell-ECM interactions.
