Abstract
BACKGROUND: Non-animal technologies (NATs; also termed new approach methodologies, NAMs) are rapidly reshaping biomedical research, safety testing and drug development by improving human relevance and reducing reliance on animal models. In Australia, substantial expertise exists across organoids, microphysiological systems and in silico approaches; however, national visibility of capabilities, infrastructure access and system-level needs has been limited. METHODS: We conducted a cross-sectional, online baseline survey (February-October 2025; https://redcap.ohmr.health.nsw.gov.au/surveys/?s=3EWRM48PEXWLFJC7.) integrated into NAT-Net registration and implemented using REDCap. The survey captured organisational characteristics, roles within the NAT ecosystem, research domains, model types, development stage, supporting services and infrastructure access, engagement preferences, and perceived challenges. Quantitative data were summarised descriptively, and free-text responses were analysed thematically. RESULTS: Of 135 submitted records, incomplete entries (n = 2) and duplicates (n = 9) were removed, yielding 124 unique respondents. Respondents were predominantly based in New South Wales (71.0%), followed by Victoria (18.5%) and Queensland (7.3%). Most organisations reported involvement in research and development (79.0%), education and training (41.9%), ethics and the 3Rs (39.5%), and provision of supporting services or facilities (32.3%). Among respondents engaged in NAT R&D (n = 102), commonly reported approaches included in vitro 3D models (64.7%), in vitro 2D culture (56.9%), microphysiological systems/organ-on-chip platforms (35.3%), and in silico methods (30.4%). Among service providers answering the access item (n = 40), 60.0% offered external access. The most frequently identified challenges were the absence of standards and validation processes (75.8%), limited access to resources and infrastructure (74.2%), and difficulty in establishing collaborations or partnerships (71.7%). CONCLUSION: This NAT-Net baseline survey provides an initial cross-sector ecosystem mapping centred on NAT-Net participants, revealing broad cross-sectoral activity alongside persistent system-level barriers to standardisation, infrastructure access and coordination. These findings provide a critical evidence base to inform coordinated national strategy, investment, policy development and longitudinal monitoring.