The prevalence of unfavorable lifestyle habits is leading to a growing incidence of liver disease, increasing the need for transplantation or repair of damaged tissue. Decellularization of natural tissue is a promising method to maintain a three-dimensional network for tissue regeneration, as the natural structure and components are preserved, which is an essential factor for cell survival. It is of utmost importance to develop a decellularization method that removes the cells well enough and still preserves the components of the extracellular matrix. Two detergents have been used for decellularization of liver tissue – sodium laureth sulfate (SLES) and sodium lauryl sulfate (SDS). In the decellularization protocols, the influence of medium exchange time and detergent concentration was investigated and additional ultrasound treatment was included for 1, 2, 4, and 8 hours. After decellularization, the biological components in the samples were quantified: DNA, collagen, and glycosaminoglycans. After decellularization, the samples were subjected to a digestion process and freeze-dried. After decellularization and digestion, the samples were subjected to differential scanning calorimetry and the values of normalized enthalpy and initial and maximum denaturation temperatures were determined. The results obtained show that the sample SDS 1a, treated with sodium lauryl sulfate at a concentration of 1% with a medium change of 30 minutes, is the most suitable for further testing for application in 3D printing. Despite evidence of a relationship between enthalpy and denaturation temperatures with collagen content, no direct relationship was found in the measurements, which could be due to the presence of substances other than collagen.