Conventional hydrogels used as organoid cultivation matrices (e.g. Matrigel®) contain animal-derived components in varying quantities, implicating low reproducibility of experiments and limited applicability for clinical use. Based on the strong need for developing novel, well-defined, and animal-free hydrogels for 3D cell cultures, this study presents a comprehensive analysis of pancreas organoid cultivation in two synthetic hydrogels. Besides established visualization techniques to monitor organoid formation and growth, confocal Raman microscopy was used for the first time to evaluate the gel matrices and organoid formation within the gels. The approach revealed so far not accessible information about material-cell interactions and the composition of the organoid lumen in a non-invasive and label-free manner. Confocal Raman microscopy thereby enabled a systematic characterization of different hydrogels with respect to cell culture compatibility and allowed for the rational selection of a hydrogel formulation to serve as a synthetic and fully defined alternative to animal-derived cultivation matrices.
Non-invasive analysis of pancreas organoids in synthetic hydrogels defines material-cell interactions and luminal composition
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