Cultivation And Differentiation Of Retinal Neural Stem Cells In Neonatal Rats

Due to the deficiency of the regeneration in the mature neurons, central nervous system has poor self-repair capability after injury. The achievement made in the research of neural stem cells had provid a promising treatment to the patiens with the injury or degeneration of central nervous system. The major pathologic change of glaucoma or age-related macular degeneration (ARMD), the majority and irreversibility causing-blindness oculopathy, is with the death of a great quantity neurons, there is not any effective way to treat them in clinic yet. To provid a theory base for the clinical application of retinal neural stem cell (RNSC) in the treatment of the glaucoma or ARMD, retinal neural stem cells were isolated from the neonatal rat eye retina, and to be study the induction and differentiation of the cells. Cell colonies with ability of proliferation were isolated from the retina of neonatal rat. Then BDNF or/and serum were used to induce them to differentiate. The undifferentiated cells and differentiated cells were identified by immunochemistry with specific antibodies. Results show: 1. Cell colonies are of ability of proliferation demonstrated by Brdu-ir, and show nestin ( the specific marker of neural stem cells). 2. Immunochemistry result show Map-2, NF-200 and GFAP positive cells in differentied cells and show few GAD, TH, ChAT, GABARA and NR1 positive cells in differentiated neuron-like cells. 3. Without BDNF, RNSCs have obviously cell death in long-time non-serum culture; Co-operation of BDNF and serum increases the ratio of neuron in differentiated cells. There are retinal neural stem cells in neonatal rats, and were able to differentiate into glia and neurons with the expression of some kinds of neurotransmittes or their receptors. BDNF as a kind of assistant inducer, may help RNSC to survive, differentiate, mature, and probably induce them to differentiate into neurons.