(277e) The Impact of Decellularization Agents on Renal Tissue Extracellular Matrix

Nafiseh Poornejad, Lara B. Schaumann, Travis Neuberger, Sarah Chambers, Beverly L. Roeder, Alonzo D. Cook

Brigham Young University, Provo, Utah

The combination of patient-specific cells with scaffolds obtained from natural sources could result in improved regeneration of human tissues. Decellularization of the native tissue is the first step in this technology. Effective decellularization uses agents that lyse cells and remove all cellular materials, leaving intact collagenous extracellular matrices (ECMs). Although complete removal of cellular remnants should be achieved to prevent the induction of an immune response, preserving the underlying structure and basement membrane components are of crucial importance to retain regenerative potential of a decellularized scaffold. Consequently, enough care should be taken in choosing cell-lysing agents and the procedure should be optimized for different tissues.

In this study, the impact of five decellularization agents (0.1 N NaOH, 1% peracetic acid, 3% Triton X-100, 1% sodium dodecyl sulfate (SDS), and 0.05% trypsin/EDTA) on renal tissue was examined to comprehensively compare a broad range of agents including acidic/basic solution, ionic/non-ionic detergents, and protease enzyme. The NaOH solution induced the most efficient cell removal (>90% DNA removal, p-value<0.05), and resulted in the highest amount of human renal cell viability and proliferation after recellularization, although it also produced the most significant damage to collagenous fiber networks, glycosaminoglycans (GAGs), and fibroblast growth factor (FGF). The SDS solution led to less severe damage to the ECM structure, but it resulted in less proliferation of reseeded cells. Peracetic acid and Triton X-100 preserved ECM components intact. However, these agents alone were not efficient in removing cellular materials; especially peracetic acid, which left more than 80% of cellular remnants within the ECM. Trypsin enzyme didnâ??t eliminate cellular materials and resulted in significant damage to the collagenous structure of ECM.

In summary, we suggest NaOH as the most efficient agent to remove cellular materials combined with non-ionic Triton X-100 as an optimum procedure for decellularization of renal tissue.