RGD peptide sequence is an effective cell recognition motif and used to enhance the cell adhesion on desired solid material for cell immobilization. We have synthesized CRGD, CRGD-multiple-armed peptide (MAP), RGD-MAP-C and evaluated their comparative efficacy for cell immobilization. Each peptide was assembled on gold surface and investigated by the atomic force microscopy (AFM) technique in the contact mode. The viability of immobilized animal cells was examined by an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Our results showed that RGD-MAP-C in comparison to others was the most effective proliferation of cells on the gold surface. The goal of this present work is integration to the nano-pattern cell chip bioplatform for biomedical assays or provide valuable insights into cell biology and design of biomaterials. This RGD-MAP-C can be applicable to the nano-pattern cell chip platform.