Endothelial colony-forming cells for preparing prevascular three-dimensional cell-dense tissues using cell-sheet...

Area of Science:

• Biomaterials Science
• Regenerative Medicine
• Vascular Biology

Background:

• Prefabricating vascular networks in 3D tissue constructs aids functional anastomosis with host vasculature.
• Clinical use of vascular-derived endothelial cells (ECs) is limited by invasive biopsies and low cell yield.
• Endothelial colony-forming cells (ECFCs), a blood-derived progenitor cell type, possess significant proliferative and vasculogenic potential.

Purpose of the Study:

• To fabricate prevascularized 3D cell-dense tissue constructs using cord blood-derived ECFCs.
• To evaluate the in vivo angiogenic potential of these ECFC-based constructs.
• To compare ECFCs with human umbilical vascular endothelial cells (HUVECs) in construct fabrication.

Main Methods:

• ECFCs and HUVECs were sandwiched between human dermal fibroblast (FB) sheets using a fibrin-coated cell-sheet manipulator.
• Constructs were cultured for 3 days to assess network and lumen formation.
• ECFC-containing constructs were transplanted into immune-deficient rats for in vivo evaluation.

Main Results:

• ECFCs formed vascular network structures within double-layered FB sheets, comparable to HUVECs.
• Triple-layered FB sheet constructs with ECFCs developed lumen structures after 3 days of co-culture.
• Transplanted ECFC constructs showed ECFC-lined functional microvessels containing host erythrocytes one week post-transplantation.

Conclusions:

• Cord blood-derived ECFCs can be utilized to fabricate prevascularized 3D cell-dense tissue constructs.
• ECFCs demonstrate potent in vivo angiogenic potential, forming functional microvessels after transplantation.
• ECFCs represent a viable alternative cell source for clinical applications in tissue engineering requiring prevascularized constructs.