RGD peptide sequence is an effective cell recognition motif and used to enhance the cell adhesion on desired solid material for cell immobilization. CRGD-MAP (Multiple-Armed Peptide) and RGD-MAP-C are synthesized, and the effect of these peptide sequence on cell immobilization is investigated. Each peptide sequence was self-assembled on gold surface due to cysteine residue, and the morphology of adsorbed surface was investigated by AFM observation in semi-contact mode. The immobilization of mouse embryonic stem cells was done and the viability after immobilization was examined by hemocytometer. It was observed that RGD-MAP-C was more effective peptide sequence for proliferation of stem cells on the gold surface comparing with CRGD-MAP. The proposed RGD-MAP-C can be used to the fabrication of stem cell chip platform by integration with nanopatterned surface for biomedical assays and design of drugs.