γ-Irradiated cord blood MNCs: different paracrine effects on mature and progenitor endothelial cells.
Cell-based therapies have been employed to promote neovascularization mainly through the release of paracrine factors inhibiting apoptosis and supporting migration and proliferation of resident differentiated cells. We tested in vitro pro-angiogenic effects of apoptotic cord blood-derived mononuclear cells (CB-MNCs) and their conditioned medium (CM) on mature endothelial cells (HUVECs) and peripheral blood-derived endothelial progenitor cells (ECFCs). CB-MNCs were γ-irradiated to induce apoptosis and cultured for 72 h to obtain the release of CM. MNCs viability, evaluated by flow cytometry, decreased progressively after γ-irradiation reaching 41% at 72 h. γ-Irradiated MNCs (γMNCs) released increasing amounts of EGF, PDGF-AB and VEGF in their CM over time, as assessed by ELISA. γ-MNCs and their CM enhanced capillary-like network formation (in a dose-dependent and time-persistent manner), proliferation and migration of HUVECs in vitro, while they primed capillary-like network formation (dose-independent and not time-persistent) and induced migration but did not support proliferation of ECFCs. Our data support the hypothesis of paracrine mechanism as prevalent in regenerative medicine and demonstrate the efficacy of MNCs secretome in inducing neovascularization. To our knowledge, this is the first paper highlighting differential pro-angiogenic effects of CM on mature and progenitor endothelial cells, adding a tile in the understanding of mechanisms involved in neovascularization.
Immunohematology and Transfusion Service, IRCCS Policlinico S. Matteo Foundation, Pavia, Italy. Electronic address: m.cervio@smatteo.pv.it.
Marila
M
Cervio
Service of Biometry and Statistics, Scientific Direction, IRCCS Policlinico S. Matteo Foundation, Pavia, Italy.
Luigia
L
Scudeller
Immunohematology and Transfusion Service, IRCCS Policlinico S. Matteo Foundation, Pavia, Italy.
Gianluca
G
Viarengo
Research Center for Regenerative Medicine, IRCCS Policlinico S. Matteo Foundation, Pavia, Italy.
Manuela
M
Monti
Immunohematology and Transfusion Service, IRCCS Policlinico S. Matteo Foundation, Pavia, Italy.
Claudia
C
Del Fante
Vascular Surgery Unit, IRCCS Policlinico S. Matteo Foundation, Pavia, Italy.
Vittorio
V
Arici
Immunohematology and Transfusion Service, IRCCS Policlinico S. Matteo Foundation, Pavia, Italy.
Cesare
C
Perotti
D017209:Apoptosis; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D017077:Culture Media, Conditioned / Q000737:chemistry; D042783:Endothelial Cells / Q000528:radiation effects*; D004797:Enzyme-Linked Immunosorbent Assay; D005312:Fetal Blood / Q000528:radiation effects*; D005720:Gamma Rays; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology*; D009389:Neovascularization, Pathologic; D010641:Phenotype; D012038:Regeneration; D013234:Stem Cells / Q000166:cytology*; D013997:Time Factors
D017077:Culture Media, Conditioned
Conditioned medium; Cord blood; Endothelial cells; Endothelial progenitor cells; Mononuclear cells; Neovascularization