Overview
This video describes the preparation of a decellularized rat kidney. The obtained acellular vascular scaffold can serve as a promising platform for tissue engineering of organ grafts.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Kidney Harvesting and Decellularization
- Harvest the left kidney with the abdominal aorta and inferior vena cava.
- Ligate the ureter, thoracic aorta, superior vena cava, and branches of the abdominal aorta.
- Keep the organ hydrated in Dulbecco's PBS (DPBS) in a 10 cm Petri dish.
- Cannulate the abdominal aorta and inferior vena cava with a 23G catheter. To remove residual blood, connect the cannula with a peristaltic pump and wash with DPBS (500 mL) and 16 U/mL heparin for 90 min at a rate of 5 rpm at 37 °C.
- To decellularize the kidney, perfuse the kidney with 1% Triton X-100 (1 L) for 3 h and then with 0.75% sodium dodecyl sulfate (SDS) solution (2 L) for 6 h at a constant pressure of 40 mmHg.
NOTE: The kidney will become transparent after 8 h. - To remove residual SDS, perfuse the sample with 1% penicillin in distilled water (6 L) for 18 h (overnight) and then with sterile DPBS (500 mL) and 16 U/mL heparin for 90 min.
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Materials
| Name | Company | Catalog Number | Comments |
| Polyethelyne 50 tubing, catheter tubing 100 ft | Braintree Scientific | .023" × .038” | |
| 3-0 silk spool vascular access/ ligation in rat | Braintree Scientific | SUT-S 110 | |
| Fine scissors to cut fascia/ connective tissue | Fine Science Tools | 14058-09 | |
| 25 gauge inch guide needle(for vascular catheters) | Becton Dickinson | 305145 | |
| Schwartz microserrefine vascular clamps | Fine Science Tools | 18052-01 (straight) | |
| 5-0 silk spool vascular access/ ligation in mouse | Braintree Scientific | SUT-S 106 |
