The spiny mouse (Acomys) is gaining popularity as a research organism due to its phenomenal regenerative capabilities.Acomysrecovers from injuries to several organs without fibrosis. For example,Acomysheals full thickness skin injuries with rapid re-epithelialization of the wound and regeneration of hair follicles, sebaceous glands, erector pili muscles, adipocytes, and dermis without scarring. Understanding mechanisms ofAcomysregeneration may uncover potential therapeutics for wound healing in humans. However, access toAcomyscolonies is limited and primary fibroblasts can only be maintained in culture for a limited time. To address these obstacles, we generated immortalizedAcomysdermal fibroblast cell lines using two methods: transfection with the SV40 large T antigen and spontaneous immortalization. The two cell lines (AcoSV40 and AcoSI-1) maintained the morphological and functional characteristics of primaryAcomysfibroblasts, including maintenance of key fibroblast markers and ECM deposition. The availability of these cells will lower the barrier to working withAcomysas a model research organism, increasing the pace at which new discoveries to promote regeneration in humans can be made.