Bone Marrow Stromal Cells Cultured In Vitro And The Study Of Transplanted Into The Vitreous Cavity

Objective Retina is an important part of the optic nerve system. The retinal diseases are common in clinic, which can lead to blindness, such as glaucoma,retinitis pigmentosa(RP), optic nerve trauma and age-related macular degeneration(AMD). But the therapeoutic result was not contented for the injured retina, especially for the serious damage and the disease of an advanced stage. Only acquiring plenty of cells which have the characters of the retinal cells can we cue these diseases. With the deepen development of stem cell reseach, It is possible to adopt the cell substitution therapy. In this study we cultured bone marrow stromal cells in vitro and transplanted them into the rabbit vitreous body to observe the survival of BMSCs after transplanted into vitreous cavity, in order to finding a good transplanting way and possible orginal cells for the therapy of the retinal diseases.Methed:The BMSCs from rabbit were isolated by its chsrscteristic of growing adherence to the plastic surface with cell culture, rabbit bone marrow, taken from tibias, were plated in DMEM medium. Through the change of medium and passage, HSCs were removed. BMSCs were passaged by the rate of 1:2. The third passage cells were labeled using DAPI. After digestion and centrifugation, the cells were disseminated in DMEM medium without serium about the density of 5 X 106/ml. O.lml of the cells suspension were slowly injected into the intravitreal space with the injector. After injection the fudus were observed every week. 4 weeks later rabbits were sacrificed, eyeballs were enucleated for continous frozen slides. HE staining and DAPI fluorescence are used to identify the transplanted cells of bone marrow origin.Result: BMSCs were adherent to the plastic surface of the cell culture dish and growed fast 3 day later. The morphology of BMSCs tended homogenous after 7-10 days. Cells had fibroblastic-like morphology. The passage cells had the same morphology. HE staining shows there are cell masses in vitreous cavity. These cells are DAPI labled cells under fluorescence microscopy.Conclusion: BMSCs were cultured easily and explanded quickly in vitro. We can acquire BMSCs by its characteristic of growing adherence to the plastic surface with cell culture. BMSCs can be cultured and passaged for long time in vitro with the same morphology. The transplanted cells can survive in vitreous cavity for 4 weeks at least and intend to differentied into retinal cells. Transplanting into vitreous cavity is a simple and possible way.