02059nas a2200229   4500000000100000000000100001008004100002260000900043653002000052653002500072653001100097653001300108653002500121100001500146700001700161245008800178856006700266300001100333490000700344520146400351022001401815       2025                            d  c202510a3D cell culture10aextracellular matrix10amatrix10aorganoid10atranslational models1 aLisa Wolff1 aSven Hendrix00aRethinking Matrigel: The Complex Journey to Matrix Alternatives in Organoid Culture  uhttps://onlinelibrary.wiley.com/doi/abs/10.1002/advs.202508734  ae087340 v123 aMatrigel is a crucial tool in cell biology, particularly for organoid research. It forms a stable scaffold with components like basement membrane proteins and growth factors, resembling the extracellular matrix and mimicking the cellular microenvironment. Its complex composition is both an asset and a drawback, as it is undefined and can vary from batch to batch. Another issue is the murine origin of Matrigel, raising ethical and scientific concerns. Interspecies variation hinders the successful translation of research findings from experimental models to clinical application in humans. Despite these well-known and often-discussed disadvantages, Matrigel is often the first choice of matrix. This review explores why Matrigel remains the gold standard for many human 3D culture systems despite its murine origin and well-known limitations. Therefore, challenges are identified that prevent researchers from transitioning to matrix alternatives and, eventually, developing completely xeno-free human model systems. To tackle these challenges, it is suggested to move beyond a one-for-all approach by pursuing a tissue- and model-specific focus when designing new matrices or selecting an alternative from already available options. To facilitate the implementation of Matrigel substitutes, we provide a matrix selection checklist and a scaffold assessment tool, and quantitative assessment criteria to evaluate matrix-model compatibility are suggested.  a2198-3844