Marine wildlife species, such as whales, dolphins, sharks, sea turtles and dugong, play critical roles in the structure and function of marine ecosystems. However, the biology of marine wildlife remains poorly understood, primarily due to the logistical and ethical constraints on conducting research on these large, long-lived, and often threatened, species. Cell cultures are an increasingly valuable, ethical and novel tool for research into biological processes and health assessment of humans and animals. However, their application to understanding the biology and health of marine wildlife has been limited, largely driven by the lack of available cell cultures for these species. In collaboration with marine wildlife rehabilitation centres, we have established and validated cell cultures from a range of marine wildlife species (whales, dolphins and turtles) and tissue types (skin, liver kidney, brain). Further, we have characterised these cell cultures by confirming the species of origin, and developed a platform for distributing these cell cultures to researchers globally. Cell cultures from the Marine Wildlife Cell Bank are now facilitating species-specific understanding of wildlife biology and ecology, contributing to the management and conservation of marine wildlife in Australia.