Human umbilical cord blood plasma can replace fetal bovine serum for in vitro expansion of functional human endothelial colony-forming cells.
OBJECTIVE
A hierarchy of endothelial colony-forming cells (ECFC) with different levels of proliferative potential has been identified in human circulating blood and blood vessels. ECFC has recently become an attractive target for new vascular regenerative therapies; however, in vitro expansion of ECFC typically depends on the presence of fetal bovine serum (FBS) or fetal calf serum (FCS) in the culture medium, which is not appropriate for its therapeutic application.


METHODS
To identify optimal conditions for in vitro expansion of ECFC, the effects of human endothelial serum-free medium (SFM) supplemented with six pro-angiogenic cytokines and human umbilical cord blood plasma (HCP) were investigated. The in vitro morphology, proliferation, surface antigen expression and in vivo vessel-forming ability were utilized for examining the effects of medium on ECFC.


RESULTS
This novel formulation of endothelial cell culture medium allows us, for the first time, to isolate and expand human ECFC efficiently in vitro with a low concentration of HCP (1.5%) and without bovine serum additives. In this serum-reduced medium (SRM), human ECFC colony yields remained quantitatively similar to those cultured in a high concentration (10%) of bovine serum-supplemented medium. SRM-cultured ECFC displayed a robust clonal proliferative ability in vitro and human vessel-forming capacity in vivo.


CONCLUSIONS
The present study provides a novel method for the expansion of human ECFC in vitro and will help to advance approaches for using the cells in human therapeutic trials.
Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana 46202, USA.
Lan
L
Huang
Paul J
PJ
Critser
Brenda R
BR
Grimes
Mervin C
MC
Yoder
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology*; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects; D005312:Fetal Blood / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D016130:Immunophenotyping
D003470:Culture Media; D016895:Culture Media, Serum-Free
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