This study examined the effects of the surface properties of the materials, such as the hydroxyl, methyl and amino groups, on rat bone marrow derived Mesenchymal Stem Cell (MSC) seeding. A series of hydrogels were prepared in film form using 2-hydroxyethyl methacrylate (pHEMA), poly(ethyleneglycol) methacrylate (PEG-MA), and/or hydroxypropyl-chitosan (HPC). The physicochemical properties of these hydrogel films, such as water content, functional groups, contact angle, surface energy and thermal properties were affected by the composition of the materials. The ability of the MSCs to form colonies, as well as their viability on these materials were also analyzed. The water content of the hydrogel films increased with increasing PEG-MA and HPC ratio in the hydrogel. Contact angle measurements of the surface of the hydrogel films demonstrated that all the materials gave rise to a significantly hydrophilic surface compared to pure pHEMA. The blood protein interactions and platelet adhesion were reduced significantly on the surface of the materials upon the incorporation of PEG-MA compared to the control pure pHEMA and vice versa for HPC. The ability of the MSCs to adhere and form colonies on these materials was also analyzed. The results showed that these materials are suitable candidates to isolate and expand MSCs.