Cryostructuring of polymer systems. 47. Preparation of wide porous gelatin-based cryostructurates in sterilizing organic media and assessment of the suitability of thus formed matrices as spongy scaffolds for 3D cell culturing

New gelatin-based cryostructurates have been elaborated and tested as scaffolds for three-dimensional (3D) cell culturing. Scaffold preparation included dissolution of Type A gelatin in dimethylsulfoxide, freezing of such solution, cryoextraction of crystalline phase with cold ethanol, cross-linking of gelatin with carbodiimide in ethanol medium, treatment of the matrix with ethanolic solution of Tris and tanning of the matrix with formaldehyde dissolved in ethanol. The use of organic media during all the preparation stages ensured the sterility of the scaffolds. The matrices thus prepared were seeded with human adipose tissue multipotent mesenchymal stromal cells to confirm the biocompatibility of scaffolds and their possibility to provide necessary environment for the cell growth and differentiation. The cells attached onto the surface of the pore walls, proliferated and differentiated into osteogenic and adipogenic lineages. These results demonstrate that gelatin-based cryostructurates prepared in the sterility ensuring organic media can be used as scaffolds for tissue engineering purposes.