Purpose In this study, differentiation properties of optic cup-derived retinal stem cells (OC-RSCs) towards ganglion cells were analyzed.
Methods OC-RSCs were isolated from optic cups of embryos at embryonic day ED 12.5 (tailbud stage) of Long Evans rat, the expression of Thy1.1 was estimated by immunochemistry and western blot, the number of Thy1.1 postive cells was detected by flow cytometry, after treatment with a series of concentrations of FBS in the culture medium.
Results Addition of FBS led to the production of Thy1.1-positive cells, but the differentiation potential was affected by FBS concentration. After 21 days, 91.90% of cells expressed Thy1.1 was produced with 0.5% FBS in the culture medium, and 57.65% in 2.5% FBS, 82.18% in 5% FBS, 84.26% of cells with the addition of 10% FBS. But the appearance of Thy1.1+ cells, which have larger, irregular phase-bright cell bodies as spindle-shaped cells, were different from that of ganglion cells cultured in vitro.
Conclusion OC-RSCs preferentially generate ganglion cells as depicted by the expression of Thy1.1. The differentiation towards ganglion-like cells was FBS concentration-dependent, and the highest percentage of Thy1.1-positive cells was found using 0.5% FBS. However, the Thy1.1+ cells do not have the morphological properties of typical cultured ganglion cells in vitro, it was the potential explanation that the condition for induction were not suitable for growth and maturation of ganglion cells.
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