Streamlined System for Conducting In Vitro Studies Using Decellularized Kidney Scaffolds

Abstract

Kidney regeneration is a complex process that can only be studied in vitro at a limited capacity due to the inherent structural and functional complexity of its tissues. Thus, a suitable platform for conducting cellular response and development should be established from decellularized tissues with intact microarchitecture. In this study, a modular streamlined system was developed to allow manageable handling and setup of in vitro studies using decellularized rat kidneys. The system is composed of commercially available parts that can be reused, interchanged, and reconfigured based on the desired experimental stage and process. Decellularization was confirmed through time-lapse observation, stained tissue sections, genetic material quantification, and protein analysis. The capacity of the bioreactor design to support cell-seeded decellularized kidney constructs was tested by determining viability of seeded podocytes and endothelial cells. Based on the results, decellularized kidneys with renal proteins and intact microstructures can be achieved in relatively shorter periods compared (12h) to established protocols (96-120h). The minimalistic kidney bioreactor design not only maintained sterility of decellularized kidney without cells but also permitted manageable maintenance of cell-immobilized constructs for up to 1 week. Through this streamlined system, sustainable and reproducible in vitro experiments for kidney regeneration can be designed and conducted using decellularized kidney as a platform for cell growth and development.