Initial attachment and spreading of the inoculated cells determines the long-time viability of cells onto biomedical scaffolds designed for various orthopedic or other clinical applications. The aim of this study was to investigate the influence of biomimetic thin film coating surfaces of bio-derived bone scaffolds with collagen proteins and chitosan on bone marrow mesenchymal stem cells interactions in order to improve cell adhesion, spreading and proliferation. These two merits were used synthetically to generate apatite-based materials that can function as allograft bone grafts in humans. In this study, the thin film coatings were operated by means of soaked, pre-frozen, and freeze-dried step by step. All coatings were characterized using Raman spectra, inverted microscope, atomic force microscopy, and scanning electron microscopy. After that, the bio-derived bone scaffolds with or without thin film coatings were used in bone marrow mesenchymal stem cell culture experiments to study cell adhesion, spreading, viability, proliferation and morphology. Then, the biological morphologies of the fabricated cell-scaffold constructs were detected by scanning electron microscope (SEM). The cell reactions were investigated concerning cell adhesion, migration, spreading, and proliferation under inverted microscope and fluorescence microscopy. The results showed that the bio-derived bone scaffold treated with thin film coatings by using rat-tail type I collagen and chitosan improved the adhesion and spreading of mesenchymal stem cells in comparison to the untreated one. Besides, cell viability and morphology were not affected by the presence of either type of thin film coating. Still, the results assay revealed an increased proliferation of bone marrow mesenchymal stem cells on both types of thin film coatings compared to coating with non-coated controls.