The hydrophilicity of the cell culture substrate was controlled by depositing it on the alumina surface through the laser-induced backward transfer (LIBT) method. Alumina particles were sized using laser energy density and deposited on the soda lime glass surface. The particle size and hydrophilicity of the alumina deposition surface were evaluated by measuring the surface roughness, contact angle, and light diffusivity. As the particle size increased, the effect of alumina became stronger, and the deposited surface had relatively higher roughness, stronger hydrophilicity, and higher light diffusivity. The stronger the alumina effect, the lower the growth of Staphylococcus aureus on the deposited surface. In this study, it was confirmed that selective bacterial growth and culture could be controlled by adjusting the strength of the alumina coating using the LIBT process.