DEVELOPMENT OF TISSUE-CUSTOMIZED DECM EXTRACTION STANDARDIZATION PROCESS.

Decellularized extracellular matrix (dECM) is a versatile functional biomaterial that has a structure and composition specific to the organ and is involved differently in the functional properties of cells, tissues and organs. The remaining cells or nucleic acid fragments must be completely removed during the decellularization process that removes cells while preserving biological properties in the tissue, which is essential because it can cause immune resistance reactions and inflammation. Methods for decellularization are diverse and widely studied, but there is still no definitive method for decellularization due to the conditions for successful decellularization and the pathological and structural differences of each organ. This study identified customized decellularization method for porcine-derived organs. Porcine-derived biological tissues were decellularized in a mixture of 0.5% SDS, 1% Triton X-100, and dECM was extracted by physical, chemical, and enzymatic methods including Pepsin. Finally, sterilization is performed by irradiating 1 kGy of electron beam. Components such as dsDNA, GAGs, and collagen were analyzed, and decellularization was evaluated by scanning electron microscope (SEM) and electrophoresis. To evaluate the safety of the cells, HT-22 and Hep G2 cells were cultured with various concentrations of dECM added to the culture medium to observe cell proliferation. The results showed that the dsDNA content in porcine-derived dECM was significantly reduced by more than 99% by keeping it below 10 ng/mg (dry weight) (p<0.0001). In addition, the content of GAG and collagen was mostly maintained, and the size of DNA fragments was smaller than 100 bp. No visible macroscopic nuclear material was observed, and at the same time, collagen fibers and elastic fibers were also identified. In cell culture, the proliferation rate of cells with dECM added to the culture medium was significantly increased. In conclusion, it has been proven that dECM can be effectively extracted through an organ-specific customized decellularization method. Furthermore, it is believed that it can be widely used as a material for cell and basic experiments using dECM through cell safety experiments. [ABSTRACT FROM AUTHOR]