Manufacturing considerations for producing and assessing decellularized extracellular matrix hydrogels

Overview of a decellularization process for skeletal muscle and accompanying experimental design considerations for the manufacturing of decellularized extracellular matrix hydrogels. [Display omitted] •Degree of animal-to-animal variability for porcine tissue source.•Introduction of a bioburden reduction step and impact on final decellularized extracellular matrix product.•Evaluation of several harvesting conditions for potential raw material sources. Although several decellularized extracellular matrix (ECM) sheets or patches have been commercialized for use in the clinic, only one injectable decellularized ECM hydrogel, a decellularized myocardial matrix, has reached clinical trials. Consequently, very little information is available for established manufacturing standards or assessments of these materials. Here we present detailed methodology for investigating three parameters related to manufacturing optimization for a porcine derived skeletal muscle ECM hydrogel – animal-to-animal variability, bioburden reduction, and harvesting conditions. Results from characterization assays, including residual dsDNA content and sulfated glycosaminoglycan content, did not yield noteworthy differences amongst individual animals or following the addition of a bioburden reducing agent. However, the tissue collected under different harvesting conditions contained varying amounts of fat, and the protein compositions of the decellularized products differed, which could ultimately impact subsequent efficacy in vitro or in vivo. As decellularized ECM hydrogels continue to be evaluated for various applications, the differences between laboratory-scale and manufacturing-scale material batches should be thoroughly considered to avoid costly and timely optimization during scale-up.