Advertisement

Vessel wall–derived endothelial cells rapidly proliferate because they contain a complete hierarchy of endothelial progenitor cells

David A. Ingram, Laura E. Mead, Daniel B. Moore, Wayne Woodard, Amy Fenoglio and Mervin C. Yoder

Article Figures & Data

Figures

  • Figure 1.

    Immunophenotypic analysis of cord blood EPC-derived endothelial cells, HUVECs, and HAECs. Immunophenotyping of cell monolayers derived from cord blood EPCs (A), umbilical veins (B), or human aortas (C) by fluorescence cytometry. Cord blood EPC-derived ECs, HUVECs, and HAECs express CD31, CD141, CD105, CD146, CD144, VWF, and Flk-1 but do not express CD45 and CD14. Shown are representative data from 5 independent experiments using 5 different cord blood EC monolayers, 5 different HUVEC samples, and 5 different HAEC samples with similar results. Isotype controls are overlaid in white on each histogram for each surface antigen tested.

  • Figure 2.

    Quantitation of the clonogenic and proliferative potential of single cord blood endothelial cells derived from EPC colonies, HUVECs, and HAECs. (A) The percentage of single cord blood (CB) EPC-derived ECs, HUVECs, or HAECs undergoing at least one cell division after 14 days of culture. Results represent the average of 5 independent experiments using single ECs derived from different donors. (B) Number of cell progeny derived from a single CB EPC-derived EC (□), HUVEC (▦), or HAEC (□) in an individual well after 14 days of culture. *P < .01 by Student paired t test for comparison of a single CB-derived EC versus either a single HUVEC or HAEC. (C) Representative photomicrographs (× 50 magnification) of the different EC clusters (less than 50 cells) or colonies (more than 50 cells) derived from a single cord blood EPC-derived EC, HUVEC, or HAEC. Results are representative of 4 other independent experiments utilizing cells from different donors. Scale bar in photomicrographs represents 100 μm. (D) Percent of the cell progeny derived from a single cord blood EPC-derived EC, HUVEC, or HAEC that formed secondary colonies or rapidly grew to cell confluence after 7 days of culture in a 24-well tissue culture plate. Results represent the average ± SEM of 4 independent experiments using cells derived from 4 different donors. *P < .01 by Student paired t test for comparison of a single CB-derived EC versus either a single HUVEC or HAEC. (E) Representative photomicrographs (× 50 magnification) of the secondary EC colonies or confluent cell monolayers derived from the cell progeny of a single plated cord blood EPC-derived EC, HUVEC, or HAEC in a 24-well plate after 7 days in culture. Scale bar in photomicrographs represents 100 μm. (F) Telomerase activity in an HPP-ECFC (HPP) and LPP-ECFC (LPP) colony derived from HUVECs. “Pos” indicates telomerase activity in HeLa cells, which were used as a positive control, and “neg” indicates a negative control. Results are representative of 4 other independent experiments. Similar differences in telomerase activity were observed between HPP-ECFC and LPP-ECFC colonies isolated from cord blood ECs and HAECs (data not shown).