Polydopamine (PDA) is the final oxidation product of dopamine or other catecholamines. Since the first reports of PDA coatings starting around 2007, these coatings have been widely studied as a versatile and inexpensive one-step coating option for biomaterial functionalization. The coating attach to a wide range of materials and can subsequently be modified with biomolecules or nanoparticles. However, as a strong candidate for biomaterial research and even clinical use, it is important to unravel the changes in physico-chemical properties and the cell-PDA interaction as a function of heat sterilization procedures and shelf storage periods. Four groups were examined in this study: titanium (Ti), PDA-coated Ti samples and PDA-coated Ti samples either stored for up to two weeks at room temperature or heated at 121 °C for 24 h, respectively. We used X-ray Photoelectron Spectroscopy (XPS), Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) and Water contact angle (WCA) to characterize chemical composition and surface properties of the groups. Cell adhesion and proliferation was examined by three different cell types: human primary dermal fibroblasts (hDF), human epidermal keratinocytes (HaCaTs) and a murine preosteoblastic cell line (MC3T3-E1), respectively. Cells were cultured on PDA coated samples for 4 h, 3 days and 5 days. Both thermal treatment of PDA at 121℃ for 24 h and storage of the samples for 2 weeks increased the amount of quinone groups at the surface and decreased the amount of primary amine groups as detected by XPS and ToF-SIMS. Even though these surface reactions increased the WCA of the PDA coating, we found that the post-treatments increased cell proliferation for both hDFs, HaCaTs and MC3T3-E1 s as compared to pristine PDA. This emphasizes the importance of post-treatment and shelf-time for PDA coatings.