Preparation of decellularized kidney scaffolds in rats

  Background  Decellularized scaffolds based on natural kidneys can be used in bioartificial kidneys. However, there is no standard for the preparation of kidney decellularized scaffolds. Since the result of decellularization is directly related to the subsequent recellularization and the performance of kidney function, it is necessary to explore and optimize the preparation of decellularized scaffolds.

  Objective  To explore the preparation of decellularized scaffolds based on intact rat kidneys and provide bio-scaffold materials for the preparation of tissue-engineered kidneys.

  Methods  Preparation of decellularized rat kidney scaffold was done by perfusing the isolated kidney with SDS solution to elute the cellular components in the kidney. The tissue structure and extracellular mechanism retention were observed by pathological staining and transmission electron microscopy. The integrity of the scaffold and the patency of the vascular network were observed by ultrasound and contrast-enhanced ultrasound. The nucleic acid in scaffold and residues of eluent were quantitatively detected by DNA and SDS, and cytotoxicity test were used to verify the safety of the prepared decellularized scaffold.

  Results  The decellularization time decreased with the increase of the perfusion flow rate. The high flow rate perfusion eluted the kidney, which caused the area of Bowman's capsule to become larger and then ruptured, with residual nuclear components. The decellularized scaffold prepared at low flow rate had a complete structure without cell components, and the scaffold had no cytotoxicity. The structure of decellularized scaffold perfused and eluted in a liquid environment at a flow rate of 0.5 mL/min were integrity, with DNA and SDS residues meeting the standards. The decellularized scaffold was intact by ultrasound exploration, and a integrity vascular network was visible on contrast examination.

  Conclusion  The optimal flow rate for preparing rat kidney decellularized scaffold is 0.5 mL/min. Ultrasound and contrast-enhanced ultrasound can be used as a pre-examination method for renal decellularized scaffold.