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. 2020 May 15;8:309. doi: 10.3389/fcell.2020.00309

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Copyright © 2020 Farkas, Simara, Rehakova, Veverkova and Koutna.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Characterization of cells derived by our synergistic three-phase protocol and their comparison with somatic endothelial cells (ECs). Representative flow cytometry histogram overlays represent the surface expression of CD31, CD144, CD34, KDR, CXCR4, and CD157 and uptake of Dil-labeled and acetylated low-density lipoprotein (Dil-ac-LDL). (A) CBIA-50–derived ECFCs by the end of the first passage; ECFCs were obtained by passaging CBIA-50–derived cells, analyzed in Figure 2D, by day 5 of differentiation without separation. (B) Human saphenous vein EC cell line C2 by the end of passage 7; this cell line was harvested and characterized in our laboratory. (A,B) Morphology of confluent cells shows similar phenotype in both cell lineages shown. Both cell lines have similar CD31 expression but the CBIA-50–derived cells have much higher expression of markers CD34, KDR, CXCR4, and CD157 than the HSVEC line C2. (C) Uptake of Dil-ac-LDL by CBIA-50–derived cells in passage 3 was analyzed by flow cytometry, and the entire population was positive. (D) CBIA-50–derived cells formed tubes on Matrigel.