Conjunctiva mesenchymal stem cells encapsulated in fibrin gel for retinal tissue engineering

IntroductionStem cell-based therapies are steadily gaining attractions for regenerative medicine approaches to treating retinal diseases and injury to the eye. One of the limitations in stem cell therapy is stem cell death following post-injection whit preventing functional integration with retinal tissue. Fibrin gel, a bio polymeric material, provides numerous advantages as a tissue engineering scaffold and a stem cell carrier. Fibrin gel exhibits excellent biocompatibility, promotes cell attachment, and can degrade in a controllable manner.MarerialsCurrent research is focusing on developing fibrin hydrogel scaffolds to protect stem cells during delivery and to stimulate endogenous regeneration through interactions of transplanted stem cells and retinal tissue. In this study fibrin gel was used as hydrogel scaffold for immobilization of cells.The structural characteristics of fibrin gel scaffold were examined with SEM. Rheological properties of fibrin gel were measured by rheometer and biodegradation rate of fibrin was measured for 2 weeks. After isolation of stem cellsCJMSCs, the cells were differentiated into photoreceptor-like cells by exposing with taurin as induction medium for 14 days in tissue culture plate (2D group) and fibrin hydrogel (3D group). The attachment of cells was analyzed with SEM and MTT. The expression of redopsin, nestin and RPE65 as photoreceptor-like cell markers was evaluated by immunocytochemistry and quantitative real-time PCR (RT-PCR) in 2D and 3D groups.ResultsThe results of SEM analysis showed CJMSCs were well attached in all the gels and there were good integrity between cells and scaffold. There was no toxicity effect of fibrin hydrogel on cultured cells and the viability of photoreceptor cell-like cells was higher in 3D fibrin hydrogels in comparison with 2D groups. After 2 weeks, the expression of rhodopsin, nestin and RPE65 as special markers of photoreceptor cells were detected by Real time PCR and immunofluorescence that these expressions in 3D groups were higher than 2D groups.ConclusionOur findings showed that application of readily available sources of adult stem cells like human conjunctiva stem cells with fibrin gel would be a promising strategy to increase the number of photoreceptor progenitor cells for repair and regeneration of retina disease such as photoreceptor injury

کلیدواژه ها:

Retina tissue engineering ، Photoreceptor ، Fibrin hydrogel ، 3D culture