Matrix (scaffold, matrix, framework, template) is a bioresorbable or non-bioresorbable material that can be filled with stem or somatic cells in/ex vivo in order to obtain a tissue-engineering structure for restoration of a lost organ, part of an organ, tissue. Scaffold must be to the extent necessary strong, non-immunogenic, bioactive. The porosity of the matrix must be open, the surface is rough and, most importantly, the matrix must contain factors of chemotaxis of endo- or exogenous origin, cell adhesion of their proliferation, differentiation. In this context, on the example of creating a decellularized small intestine matrix, a number of fundamental issues are highlighted regarding the choice of matrix material, its production technology, matrix evaluation in accordance with the criteria that correspond to the matrix for tissue engineering, and possible directions for its use. As a result, a non-immunogenic extracellular matrix of the small intestine was obtained by the method of detergent-enzymatic perfusion decellularization, which was sufficient in characteristics for use in various areas of tissue engineering, including plasty of defects of the skin, mucous membranes, small intestine, etc.