HaCaT cells in wound healing research: mechanistic insights, in vitro applications, and translational perspectives

HaCaT cell is an immortalized human keratinocyte cell line that is widely used as an in vitro model in the research field of wound repair. In this review, we provide an overview of the applications, molecular mechanisms, and translational potential of HaCaT cells in the exploration of wound healing. For clarity and methodological rigor, we performed a narrative literature review with a structured search strategy in the major scientific databases (PubMed, Scopus, Web of Science, and Google Scholar) for studies published between 2010 and 2026. Studies were included that reported the use of HaCaT cells in wound healing assays, investigated relevant molecular pathways, and passed the quality assessment. The current knowledge was presented using key findings qualitatively synthesized. Our analysis shows that HaCaT cells are widely used in cytotoxicity assessment, migration and proliferation assays, and evaluation of oxidative and inflammatory responses. Keratinocyte function during wound healing involves major signaling pathways, including MAPK/ERK, PI3K/Akt, NF-κB, and Nrf2/HO-1. Incorporation of computational (in silico) approaches can augment mechanistic understanding and target identification. However, future studies need to consider the lack of cellular diversity and genetic variability in the HaCaT cell line, using more advanced models to fully comprehend drug-skin interactions for early-phase drug development. To summarize, HaCaT cells are a feasible and informative in vitro model for wound healing studies, and their combination with computational and 3D models can increase translational success.