Enhanced recellularization by using albumin coating with roller bottle cell culture

Abstract

IntroductionThe decellularization and recellularization is a promising approach for tissue engineering and regenerative medicine. However, the decellularization process depletes important components like glycosaminoglycans (GAGs), affecting cell attachment and causing immunogenicity. Studies have explored various surface modification strategies to enhance recellularization. MethodsTo optimize the decellularization method, we employed whole kidney perfusion and slice kidney immersion/agitation techniques. The decellularized extracellular matrix (dECM) was then analyzed using hematoxylin and eosin (H&E) staining, scanning electron microscope (SEM), and DNA quantification. To enhance cell proliferation efficiency, albumin coating and rotating culture were applied. Also, we evaluated in vitro blood clot formation on the albumin-coated dECM by immersing it in blood. ResultsAfter decellularization, the unique structures of the kidney were preserved whether cellular components were removed. Subsequently, we utilized albumin coating and rotating culture for recellularization, and observed that albumin-coated dECM not only promoted high cell proliferation rates but also prevented blood clot formation. ConclusionThe albumin-coated dECM promoted cell proliferation and reduced blood clot formation in vitro. Also, dynamic culture condition using rotating culture allowed for improved cellular penetration into the dECM, leading to a conductive environment for enhanced tissue infiltration. This new approach suggests that the combined utilization of albumin coating and rotating culture conditions can improve the efficiency of recellularization.