To investigate the role of over-expression of Math5 on the retinal ganglion cell (RGC) expression patterns in retinal progenitor cells (RPCs).RPCs were cultured and then transfected by recombinant Math5 plasmid with internal ribosome entry site and enhanced green fluorescent protein (pIRES2-EGFP-Math5; group A), with pIRES2-EGFP transfected (group B) and no plasmid transfected (group C) as control. RGCs were identified by Thy1.1 immunocytochemistry methods and analyzed by Leica Qwin V3.1 system. Real-time polymerase chain reaction was used to examine the expression of Math5-associated genes at different time points during the differentiation of RPCs.It was determined that pIRES2-EGFP-Math5 could transfect RPCs, and the transfection rate was 24.68%. After plating, it was found that three different groups of RPCs could differentiate and express retina-specific markers, including RGC marker Thy1.1. The percentage breakdown of Thy1.1-positive cells was 30.85+/-6.28% in group A, 15.84+/-3.55% in group B, and 16.22+/-3.60% in group C. The differences between the three groups were statistically significant (p<0.001). Transfection by pIRES2-EGFP-Math5 could change the expression of Delta-1, Hes1, and Brn-3b.Math5 may up-regulate RGC expression patterns in RPCs and change the expression of Math5-associated genes.