Serum derived from multiple trauma patients promotes the differentiation of endothelial progenitor cells in vitro: possible role of transforming growth factor-beta1 and vascular endothelial growth factor165.
Ischemia in various organs and tissues takes place during and as a direct result of multiple trauma (MT). Bone marrow-derived endothelial progenitor cells (EPCs) are involved in neovascularization after ischemic incidences. Here, we report that serum derived from patients with MT stimulates differentiation of EPCs in vitro from peripheral blood mononuclear cells (PBMCs). EPCs were identified by DiL-Acetyl-LDL-uptake with concomitant UEA-I-lectin binding. A significant increase in EPC numbers was noted when PBMCs were cultivated for 72 h with the serum of MT patients (n = 25) obtained at 5 days. Furthermore, serum from MT patients enhanced the functional acting of EPCs to form prevascular structures in matrigel. Reverse transcription polymerase chain reaction analysis revealed gene expression of transforming growth factor (TGF)-beta1- and vascular endothelial growth factor (VEGF) receptors 1 and 2. Reverse transcription polymerase chain reaction analysis was based on further cultivated cell preparations, which contained at least 80% EPCs. Moreover, the addition of recombinant VEGF or low concentrations of TGF-beta increased EPC differentiation. In addition, neutralization of TGF-beta1 and of VEGF165 in MT serum using specific antibodies resulted in a significant decrease in EPC differentiation. Our data indicate that TGF-beta1 and VEGF165 play a pivotal role for EPC differentiation induced by serum of polytrauma patients.
Henrich
Dirk
D
Department of Trauma Surgery, Johann-Wolfgang-Goethe University, Frankfurt/Main, Germany. d.henrich@trauma.uni-frankfurt.de
Hahn
Patrick
P
Wahl
Matthias
M
Wilhelm
Kerstin
K
Dernbach
Elisabeth
E
Dimmeler
Stefanie
S
Marzi
Ingo
I
D015153:Blotting, Western; D002454:Cell Differentiation; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology; D003470:Culture Media; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D004727:Endothelium / Q000166:cytology*; D005434:Flow Cytometry; D006801:Humans; D007511:Ischemia; D007797:Laminin / Q000494:pharmacology; D037102:Lectins / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D009389:Neovascularization, Pathologic; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D011509:Proteoglycans / Q000494:pharmacology; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D044967:Serum / Q000166:cytology*; D012769:Shock; D013234:Stem Cells / Q000166:cytology*; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000502:physiology; D053773:Transforming Growth Factor beta1; D042461:Vascular Endothelial Growth Factor A / Q000502:physiology; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism; D014947:Wounds and Injuries; D014841:von Willebrand Factor / Q000378:metabolism
D003470:Culture Media; D004338:Drug Combinations; D007797:Laminin; D037102:Lectins; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011509:Proteoglycans; D011994:Recombinant Proteins; C508960:TGFB1 protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D014841:von Willebrand Factor; C059485:matrigel; D003094:Collagen; D040301:Vascular Endothelial Growth Factor Receptor-2