Zein molecules adsorbed on surfaces with different hydrophobicity was investigated for its feasibility of cell attachment and proliferation in order to obtain information that could be applied for spatially organizing cells. Uniform hydrophilic and hydrophobic surfaces were prepared by self-assembly MHA and ODT on gold-coated glass slides. Micro patterned surfaces were generated by micro-contact printing (µCP) of MHA on gold-coated substrates and backfilling with ODT solution. Zein uniform surfaces and zein patterns were prepared by submerging the uniform and patterned substrates in zein solution and let it self-assembly on monolayers of carboxyl and methyl groups, respectively. Thus made substrates were used to culture mouse fibroblast cells (NIH 3T3), while cell culture dish was chosen as control. It was found that both the density and aspect ratio of cells were significantly higher when cell grew on zein deposited on carboxyl surfaces than it deposited on methyl surfaces. Results indicated that zein generated a polar surfaces when interact with carboxyl groups which favorite cells to attach and proliferate, while on methyl groups zein surfaces were hydrophobic. Cells seeded on patterned zein surfaces attached and proliferated exclusively on zein-MHA lines. This preliminary study demonstrates that zein is a useful growth substrate for hosting cells and further more to control spatial orientations of cells upon adsorbing on to surfaces with opposite hydrophobicity.