In this WG, research activities will be combined and coordinated to gain a common insight in corneal tissue engineering for cornea regeneration. Unique corneal stromal mesenchymal stem cells will be used to recellularise scaffolds, being developed in WG1. Researchers will work in different laboratories and three-monthly meetings are planned to define the optimum methodologies, conditions and mechanisms required to encourage cornealisation (epithelial and stromal cell differentiation from novel corneal mesenchymal stem cells) for normal corneal regeneration will be explored and characterized.
This WG has three research objectives, namely:
1. To optimize cellularisation of scaffolds whilst maintaining essential stem cell characteristics favourable for cornealisation. Different techniques used in laboratories in different countries will be validated and standardized;2. To investigate mechanobiology of scaffold cornealisation and disseminate this expertise through the consortium;
3. To investigate mechanobiology of scaffold cornealisation to influence the research progress of the network.
Research tasks to achieve these objectives are:
• Task 2.1: To optimize cellularisation scaffolds whilst maintaining essential stem cell characteristics favourable for cornealisation. Scaffolds will be cellularised by corneal stromal mesenchymal stem cells and the effects of these topographical cues on morphology will be assessed using transmission electron microscopy (TEM). For the panel of characteristic mesenchymal, flow cytometry will be used for stem cell phenotype. Corneal stem cell markers immuno-fluorescent staining (confocal microscopy) will be used to determine the distribution of any phenotypic changes in situ, and will provide information to guide necessary modification of scaffolds in WG 1. This knowledge will lead to optimisation of culturing conditions in the different laboratories.
• Task 2.2: To investigate the mechanobiology of scaffold cornealisation. Remodeling of the scaffold will be studied using transmission electron microscopy, histological staining (eosin for collagens and periodic acid-Schiff for proteoglycans) and multiplex immune-fluorescent staining (confocal microscopy). The Action’s website will be used to show the outcome, in order to stimulate the exchange of new ideas.
• Task 2.3: To optimize scaffold cornealisation to achieve optimum normal corneal characteristics with self renewal properties. The role of 3D corneal engineering solutions (bioreactors with dynamic hydrostatic pressures) will be investigated to enhance the culture conditions (mechanical and nutritional) required to complete development of normal epithelial architecture of the cornea whilst maintaining maximum self renewal capacity for long term survival. Knowledge about culture conditions will be exchanged to optimise cell growth through mutual meetings.