Abstract:

BACKGROUND: Domestic studies on in vitro isolation and identification of retinal stem cells are still in the preliminary exploration stage.
OBJECTIVE: To isolate, culture and identify the retinal stem cells from newborn rats in vitro and to investigate the cell multi-differentiation capacity.
METHODS: The cells derived from the ciliary marginal zone (CMZ) of newborn 24-hour SD rats were isolated and cultured in vitro. The proliferating capacity and differentiating properties of the cultured cells were studied by using immunocytochemistry methods.
RESULTS AND CONCLUSION: The isolated cells derived from CMZ cultured in serum-free medinum could give rise to neural spheres in the presence of basic fibroblast growth factor and epidermal growth factor. These cells could proliferate successively and generate secondary neural spheres, thus displaying potential to self-renew. These neural spheres could be expanded for up to 8 passages and could express constantly the neuroectodermal marker Nestin and Chx-10, a retinal stem cell marker, showing the undifferentiated stem cells properties in vitro. Analysis of the differentiation potential of the cultured cells in vitro showed that the cultured cells derived from CMZ were multipotential. Upon withdrawal of basic fibroblast growth factor and epidermal growth factor, and by addition of serum, the stem cells expressed cell type specific markers corresponding to neurons and glia, such as NSE, GFAP, Opsin, PKC, and β-tubulin.