Preparation of Decellularized Rat Kidney: A Procedure to Generate Acellular Vascularized Whole-organ Scaffold from a Harvested Rat Kidney

Decellularized whole-organ scaffolds consist of a three-dimensional extracellular matrix with an intact vascular network. These scaffolds retain specific tissue composition and functional integrity and play an essential role in organ bioengineering.

To prepare a decellularized rat kidney, first, obtain a freshly harvested rat kidney along with its major arterial and venous vasculature – the abdominal aorta and inferior vena cava. Tie off the ureter, minor arterial, and venous bifurcations to prevent liquid leakage during the subsequent perfusion process.

Place the kidney in a suitable buffer to ensure organ hydration. Insert appropriately sized catheters into the major vasculatures and secure them. Connect the catheter to a peristaltic pump. Perfuse the kidney with an anticoagulant-containing buffer at an appropriate flow rate and pressure for the desired duration.

This step helps to efficiently remove blood from the organ through the venous outflow. Change the perfusate to a non-ionic surfactant to permeabilize the cellular membranes. Thereafter, perfuse the organ with an anionic surfactant to effectively solubilize the cellular and nuclear membranes, resulting in cell lysis and the release of intracellular contents.

The liberated cellular components and debris flow out of the kidney, gradually rendering it translucent. Inject the decellularized kidney with a suitable buffer containing antibiotics and anticoagulants to remove residual surfactant and any remaining debris from the organ. Antibiotics prevent the likelihood of bacterial growth within the decellularized organ.