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Immunology JoVE (Open Access) Citable · DOI

Decellularization and Recellularization Methodology for Human Saphenous Veins

DOI: 10.3791/57803-v
What you'll learn
  • ✓ Perform detergent-based decellularization of human saphenous veins
  • ✓ Recellularize decellularized veins using perfusion bioreactor systems
  • ✓ Evaluate structural and cellular integrity pre- and post-processing
  • ✓ Assess endothelial cell colonization on engineered vascular scaffolds
Protocol

Here, we describe a protocol for the saphenous vein decellularization using detergents and recellularization by perfusion of the peripheral blood and the endothelial medium.

Difficulty
advanced
Total time
~7–10 days (decellularization 2–3 days, recellularization 4–7 days)
Biosafety
BSL-1

Steps

1
Prepare decellularization setups and bioreactor

Assemble decellularization apparatus and recellularization perfusion bioreactor systems according to protocol specifications. Ensure sterility and proper fluid flow configuration for downstream processing.

▶ 00:43
2
Decellularize saphenous vein using detergents

Expose human saphenous vein tissue to detergent solutions to remove cellular components while preserving extracellular matrix structure. Monitor progress and validate decellularization efficacy.

▶ 03:55
3
Recellularize decellularized vein by perfusion

Seed decellularized vein with peripheral blood cells and endothelial medium using perfusion bioreactor. Maintain optimal culture conditions to promote cell adhesion and proliferation.

▶ 06:14
4
Evaluate normal, decellularized, and recellularized veins

Compare structural integrity, cellularity, and endothelial cell coverage across untreated, decellularized, and recellularized samples using histological and immunological assessment methods.

▶ 09:22
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