Michael Boyer, MD, Laurace E. Townsend, PhD, L

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Presentation transcript:

Isolation of endothelial cells and their progenitor cells from human peripheral blood Michael Boyer, MD, Laurace E. Townsend, PhD, L.Michelle Vogel, MD, Jeffrey Falk, MD, Darlene Reitz-Vick, BS, Katrina T. Trevor, PhD, Mario Villalba, MS, Phillip J. Bendick, PhD, John L. Glover, MD Journal of Vascular Surgery Volume 31, Issue 1, Pages 181-189 (January 2000) DOI: 10.1016/S0741-5214(00)70080-2 Copyright © 2000 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Human umbilical cord monocyte cultures: A , early EC colonies (3-15 cells, arrows ) with positive staining for factor VIII (red ), VEGF-R (blue-black ) staining positively on some surrounding cells (original magnification, ×100); B , later EC colony with positive staining for factor VIII (original magnification, ×40); C , EC staining for CD31 (red; arrows ) visible on periphery of cells (original magnification, ×200); and D , EC colony identified by uptake of DiI-Ac-LDL (orange fluorescent ; original magnification, ×100). Journal of Vascular Surgery 2000 31, 181-189DOI: (10.1016/S0741-5214(00)70080-2) Copyright © 2000 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Human adult peripheral monocyte culture (gradient separation only) demonstrates line formation (arrows) that resembles vasculogenesis. (Unstained; original magnification, ×100.) Journal of Vascular Surgery 2000 31, 181-189DOI: (10.1016/S0741-5214(00)70080-2) Copyright © 2000 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Human peripheral blood cultures (magnetic bead separation): A , CD34+ cell fraction shows colony with typical EC morphologic features (unstained; original magnification, ×100); B , CD34 staining of initially CD34+ ECs, note random staining pattern (brown, arrows; original magnification, ×200); and C , factor VIII staining (red, granular ) of CD34+ EC (original magnification, ×200). Journal of Vascular Surgery 2000 31, 181-189DOI: (10.1016/S0741-5214(00)70080-2) Copyright © 2000 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 4 Overexpression of urokinase by genetically modified peripheral blood–derived endothelial cells (A) and umbilical cord–derived endothelial cells (B) . Urokinase production quantified by enzyme-linked immunosorbent assay of culture supernatants over 10 and 12 days after transduction with a retroviral vector containing the gene for human prourokinase. Control medium is culture supernatants from matched nontransduced cells. Journal of Vascular Surgery 2000 31, 181-189DOI: (10.1016/S0741-5214(00)70080-2) Copyright © 2000 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions