Advanced Animal Replacement Testing Strategies Using Stem Cell and Organoids.

Abstract

The increasing ethical concerns and regulatory restrictions surrounding animal testing have accelerated the development of advanced in vitro models that more accurately replicate human physiology. Among these, stem cell-based systems and organoids have emerged as revolutionary tools, providing ethical, scalable, and physiologically relevant alternatives. This review explores the key trends and driving factors behind the adoption of these models, such as technological advancements, the principles of the 3Rs (Replacement, Reduction, and Refinement), and growing regulatory support from agencies like the OECD and FDA. It also delves into the development and application of various model systems, including 3D reconstructed tissues, induced pluripotent stem cell-derived cells, and microphysiological systems, highlighting their potential to replace animal models in toxicity evaluation, disease modeling, and drug development. A critical aspect of implementing these models is ensuring robust quality control protocols to enhance reproducibility and standardization, which is necessary for gaining regulatory acceptance. Additionally, we discuss advanced strategies for assessing toxicity and efficacy, focusing on organ-specific evaluation methods and applications in diverse fields such as pharmaceuticals, cosmetics, and food safety. Despite existing challenges related to scalability, standardization, and regulatory alignment, these innovative models represent a transformative step towards reducing animal use and improving the relevance and reliability of preclinical testing outcomes.