Abstract
Vessel wall-derived somatic endothelial colony-forming progenitor cells (ECFCs) are key players in vascular homeostasis and regeneration. Due to their robust proliferative potential and profound vessel-forming capacity, ECFCs are considered to represent an attractive tool for vascular regenerative medicine and a promising target for antiangiogenic tumor therapy. Here, we describe an easily applicable method for isolating ECFCs directly from unmanipulated adult human blood and an animal protein-free large-scale expansion system to generate more than 100 million functional ECFCs.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Asahara T, Murohara T, Sullivan A, Silver M, van der Zee R, Li T, Witzenbichler B, Schatteman G, Isner JM (1997) Isolation of putative progenitor endothelial cells for angiogenesis. Science 275:964–967
Hill JM, Zalos G, Halcox JP, Schenke WH, Waclawiw MA, Quyyumi AA, Finkel T (2003) Circulating endothelial progenitor cells, vascular function, and cardiovascular risk. N Engl J Med 348:593–600
Case J, Mead LE, Bessler WK, Prater D, White HA, Saadatzadeh MR, Bhavsar JR, Yoder MC, Haneline LS, Ingram DA (2007) Human CD34+AC133+VEGFR-2+ cells are not endothelial progenitor cells but distinct. Primitive hematopoietic progenitors. Exp Hematol 35:1109–1118
Rohde E, Bartmann C, Schallmoser K, Reinisch A, Lanzer G, Linkesch W, Guelly C, Strunk D (2007) Immune cells mimic the morphology of endothelial progenitor colonies in vitro. Stem Cells 25:1746–1752
Yoder MC, Mead LE, Prater D, Krier TR, Mroueh KN, Li F, Krasich R, Temm CJ, Prchal JT, Ingram DA (2006) Re-defining endothelial progenitor cells via clonal analysis and hematopoietic stem/progenitor cell principals. Blood 109:1801–1809
Ingram DA, Mead LE, Tanaka H, Meade V, Fenoglio A, Mortell K, Pollok K, Ferkowicz MJ, Gilley D, Yoder MC (2004) Identification of a novel hierarchy of endothelial progenitor cells using human peripheral and umbilical cord blood. Blood 104:2752–2760
Ingram DA, Mead LE, Moore DB, Woodard W, Fenoglio A, Yoder MC (2005) Vessel wall-derived endothelial cells rapidly proliferate because they contain a complete hierarchy of endothelial progenitor cells. Blood 105:2783–2786
Melero-Martin JM, De Obaldia ME, Kang SY, Khan ZA, Yuan L, Oettgen P, Bischoff J (2008) Engineering robust and functional vascular networks in vivo with human adult and cord blood-derived progenitor cells. Circ Res 103: 194–202
Reinisch A, Strunk D (2009) Isolation and animal serum free expansion of human umbilical cord derived mesenchymal stromal cells (MSCs) and endothelial colony forming progenitor cells (ECFCs). J Vis Exp 32:1525
Au P, Tam J, Fukumura D, Jain RK (2008) Bone marrow-derived mesenchymal stem cells facilitate engineering of long-lasting functional vasculature. Blood 111:4551–4558
Schallmoser K, Bartmann C, Rohde E, Reinisch A, Kashofer K, Stadelmeyer E, Drexler C, Lanzer G, Linkesch W, Strunk D (2007) Human platelet lysate can replace fetal bovine serum for clinical-scale expansion of functional mesenchymal stromal cells. Transfusion 47:1436–1446
Schallmoser K, Strunk D (2009) Preparation of pooled human platelet lysate (pHPL) as an efficient supplement for animal serum-free human stem cell cultures. J Vis Exp 32:1523
Reinisch A, Hofmann NA, Obenauf AC, Kashofer K, Rohde E, Schallmoser K, Flicker K, Lanzer G, Linkesch W, Speicher MR, Strunk D (2009) Humanized large-scale expanded endothelial colony-forming cells function in vitro and in vivo. Blood 113:6716–6725
Hofmann NA, Reinisch A, Strunk D (2009) Isolation and large scale expansion of adult human endothelial colony forming progenitor cells. J Vis Exp 32:1524
Bartmann C, Rohde E, Schallmoser K, Purstner P, Lanzer G, Linkesch W, Strunk D (2007) Two steps to functional mesenchymal stromal cells for clinical application. Transfusion 47: 1426–1435
Acknowledgments
We are grateful to the entire team of the Stem Cell Research Unit Graz for contribution in the development of this protocol. This work has been supported by the Austrian Research Foundation (FWF) (N211-NAN], the Austrian Research Promotion Agency (FFG) (N200] grants to D.S., and The Adult Stem Cell Research Foundation (TASCRF) grants to A.R. N.A.H. and A.R are fellows of the Ph.D. program “Molecular Medicine” of the Medical University of Graz.
Author information
Authors and Affiliations
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Hofmann, N.A., Reinisch, A., Strunk, D. (2012). Endothelial Colony-Forming Progenitor Cell Isolation and Expansion. In: Singh, S. (eds) Somatic Stem Cells. Methods in Molecular Biology, vol 879. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-815-3_23
Download citation
DOI: https://doi.org/10.1007/978-1-61779-815-3_23
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-814-6
Online ISBN: 978-1-61779-815-3
eBook Packages: Springer Protocols