The Effect of Co-Culturing Human Adipose-Derived Stem Cells with Rat Retinal Cells Survival and Axonal growth

Background: Retinal nerve cells have limited regenerationability the same as the central nervous system. This regenerationdeficiency is usually attributed to the myelin structure andglial scar. If the axonal processes are impaired for any reason,their regeneration to reestablish the connection with differentparts of brain will not be possible. However, small axon-likeshoots may appear from these cells near the impaired area.There are various strategies for retinal regeneration, includinggene therapy, laser therapy and use of stem cells. Humanadipose-derived stem cells (hADSCs) are able to differentiateinto different cells and secret various neurotrophic factors thataffect other cells. So far, no study has investigated the effect ofhADSCs on retinal cells. The present study was aimed to investigatethe effect of hADSCs and their secretory environment onthe axonal survival and growth of retinal cells in vitro.Materials and Methods: The hADSCs of patients candidatefor surgery were isolated, cultured and passaged. The rat’sretina, after isolation, was placed in closed containers. Afterpreparing the supernatant of hADSCs, retina along with thecells (Retin/hADSC) as well as their supernatant (Retin/CM)with PDL/ laminin (Poly-D-Lysine/Laminin) were co-culturedfor ۲۴ hours. The survival rate was assessed by MTT and IHCmethods at days ۱, ۷ and ۱۴. The axonal growth was determinedby immunofluorescent technique and β-tub III antibody. Then,axonal growth was analyzed by ImageJ software at day ۱۴ inretinal cells.Results: In direct co-culture group (Retin+hADS), a significantdifference was found in the survival of retinal cells at day ۱۴ incomparison with day ۷ (P<۰.۰۱) and day ۱ (P<۰.۰۰۱). In indirectco-culture group (Retin+CM), a significant difference wasobserved in the survival of retinal cells at day ۱۴ compared todays ۱ and ۷ (P<۰.۰۰۱). The analysis of axonal growth in retinalcells, Auto-analysis of the number of intersections (number ofaxonal processes) and distance of axonal processes as well ascomparison of control (Retin), direct co-culture (Retin+hADSC)and indirect co-culture (Retin+CM) groups showed the numberof intersections was significantly higher in direct and indirectco-culture groups than control group (P<۰.۰۵), while the numberof intersections was significantly higher in indirect co-culturegroup than direct co-culture group (P<۰.۰۵). The results ofAuto-analysis by ImageJ software indicated the number of axonaldeformities was significantly higher in indirect co-culturegroup than control group (P<۰.۰۱).Conclusion: hADSCs along with their supernatant can increasethe axonal survival and growth in retinal cells. Given the secre tion of different factors by hADSCs and their significant effecton the nerve cells, the results of this study can help to developbasic methods for the treatment of eye diseases in the future.

کلیدواژه ها:

hADSCs ، Supernatant ، Retina ، Co-Culture