CD34/CD133 enriched bone marrow progenitor cells promote neovascularization of tissue engineered constructs in vivo

• CD34/CD133 enriched cells (EPCCs) from human BM display characteristics of mature ECs.
• Expansion of EPCCs with PL containing autologous growth factors preserves endothelial characteristics.
• EPCCs promote neovascularization in vivo.
• EPCC–MSC crosstalk is crucial for neovascularization.

Vascularization is critical for 3D tissue engineered constructs. In large size implants the ingrowth of vessels often fails. The purpose of this study was to identify an easily accessible, clinically relevant cell source able to promote neovascularization in engineered implants in vivo and to establish an autologous culture method for these cells.MSCs (mesenchymal stem cells) and an endothelial progenitor containing cell (EPCC) population were obtained from human bone marrow aspirates. The expression of endothelial-markers, uptake of acetylated low density lipoprotein (acLDL) and tube-like structure formation capability of EPCCs were analyzed after expansion in endothelial growth medium or medium supplemented with autologous platelet lysate (PL). EPCCs were co-seeded with MSCs on hydroxyapatite-containing polyurethane scaffolds and then implanted subcutaneously in nude mice.Human EPCCs displayed typical characteristics of endothelial cells including uptake of acLDL and formation of tube-like structures on Matrigel™. In vivo, EPCCs cultured with PL triggered neovascularization. MSC/EPCC interactions promoted the maturation of newly formed luminal structures, which were detected deep within the scaffold and partly perfused, demonstrating a connection with the host vascular system.We demonstrate that this population of cells, isolated in a clinically relevant manner and cultured with autologous growth factors readily promoted neovascularization in tissue engineered constructs in vivo enabling a potential translation into the clinic.

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