Fabrications of biodegradable polymer materials are of much significance in establishing living cells on artificial surfaces for tissue engineering applications. Chitosan derived by de-acetylation of chitin may be one of the important candidates for this purpose. In the present study, porous chitosan scaffolds were fabricated using freezing (-20°c to -80°c) and lyophilization (overnight) method. We established a thin film of Arg - Gly - Asp (RGD) peptide using self-assembly (SA) technique to make mechanical link between the cell and the scaffolds surface to improve its cell adhesion efficiency. The fabricated porous scaffolds were characterized by scanning electron microscopy (SEM). The pore diameters were found to be varied with different freezing temperature. The scaffolds with pore diameter of 150 µm and 100 µm (achieved by freezing at -20°c and -80°c, respectively) were subjected to cell functions study using human Mesenchymal Stem Cell (hMSC). The scaffold with 100 µm pore diameters, found to be suitable for hMSC adhesion, spreading and proliferations. Furthermore, the improved adhesion, spreading and proliferation efficiencies were reported from RGD functionalized scaffolds compared with non-functionalized one. From this study, it was concluded that the RGD functionalized chitosan scaffolds with 100 µm pore diameters established strong attachment of cells as well as mimic functions of living cells. So, the fabricated scaffold can be potentially useful tool of in vitro cellomics for tissue engineering applications.