Unlocking the potential of supercritical CO₂ for an efficient pancreatic tissue decellularization: a step towards an advanced cell therapy

Introduction: Tissue decellularization is a procedure that removes cellular and antigenic material from specific tissues, while preserving the extracellular matrix (ECM), with conditions tailored to the tissue's properties. The decellularized ECM is a promising material for cell-based therapies or for tissue defect repair through the development of scaffolds and membranes, among others. Pancreas is an organ that regulates the glucose levels in the bloodstream, primarily through insulin secretion by endocrine pancreatic β-cells. Diabetes type 2 settles on the insufficient secretion of insulin, and the consequent increase of glucose in the body. Extensive research is being conducted to find alternatives to overcome this condition, particularly on cell-based therapies, that deliver viable and functional β-cells and restore insulin production to regulate blood glucose levels effectively. Objectives: This study aims to develop and validate a decellularization protocol for porcine pancreatic tissue using Supercritical CO₂ (scCO2) technology as an innovative alternative to solvent-based processes, offering high transfer rates, diffusivity, chemical inertness, and non-toxicity [3]. Three protocols were tested: Traditional decellularization method using detergents; scCO₂-based decellularization protocol; Hybrid protocol combining both technologies. Conclusions: Results suggest that scCO₂ is the most efficient method for porcine pancreatic tissue decellularization, reducing DNA below 50 ng/mL in four cycles and achieving near-complete removal by the fifth. The traditional + scCO₂ approach also meets the threshold in four cycles but retains more residual DNA than scCO₂. This highlights scCO₂ as a faster and more effective method, fully decellularizing tissue in 20 hours, while the traditional method takes 27 hours 30 minutes and remains incomplete.