Activity of the human immortalized endothelial progenitor cell line HEPC-CB.1 supporting in vitro angiogenesis
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ORIGINAL ARTICLE
Activity of the human immortalized endothelial progenitor cell line HEPC‑CB.1 supporting in vitro angiogenesis Aneta Kantor1 · Agnieszka Krawczenko1 · Aleksandra Bielawska‑Pohl1 · Danuta Duś1 · Catherine Grillon2 · Claudine Kieda2 · Karol Charkiewicz3 · Maria Paprocka1 Received: 12 February 2020 / Accepted: 11 July 2020 © The Author(s) 2020
Abstract The human HEPC-CB.1 cell line with many characteristics of endothelial progenitor cells (EPC) was tested for its proangiogenic properties as a potentially therapeutic compound. HEPC-CB.1 cells’ potential to differentiate into endothelial cells was revealed after treating the cells with a mixture of ATRA, cAMP and VEGF, as shown by the reduced expression levels of CD133, CD271 and CD90 antigens, augmentation of CD146 and CD31, and a decrease in cell clonogenicity. The cooperation of HEPC-CB.1 with the endothelial cell line HSkMEC.2 resulted in the formation of a common network. Tube formation was significantly more effective when resulting from HEPC-CB.1 and HSkMEC.2 cell co-culture as compared to a monoculture of each cell line. The exocrine mechanism of HEPC-CB.1 and HSkMEC.2 cross talk by secreted factors was evidenced using the HEPC-CB.1 supernatant to increase the efficacy of HSkMEC.2 tube formation. The proangiogenic factors produced by HEPC-CB.1 were identified using cytokine antibody array. Out of 120 examined factors, the HEPCCB.1 cell line produced 63, some with known angiogenic activity. As in vivo the angiogenic process occurs at low oxygen tension, it was observed that in hypoxia, the production of defined factors was augmented. The presented results demonstrate that HEPC-CB.1 cells are able to both cooperate and integrate in a newly formed network and produce factors that help the network formation. The results suggest that HEPC-CB.1 cells are indeed endothelial progenitors and may prove to be an effective tool in regenerative medicine. Keywords Angiogenesis · EPC · EPC function · EPC secretome · Regeneration
Introduction Although 22 years have passed since the discovery of endothelial progenitor cells (EPC) [1], they have not yet been clearly defined. Therefore, the definition of EPC cells includes both myeloid angiogenic cells of hematopoietic Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11033-020-05662-6) contains supplementary material, which is available to authorized users. * Aneta Kantor [email protected] 1
Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Rudolfa Weigla 12, 53‑114 Wroclaw, Poland
2
Centre de Biophysique Moléculaire, CNRS UPR 4301, Rue Charles Sadron, 45071 Orléans, France
3
Medical University of Bialystok, M. Sklodowskiej‑Curie 24A, 15‑276 Bialystok, Poland
origin, expressing C D45+, and endothelial colony forming cells of mesenchymal origin, CD45 negative. The time necessary to form colonies in vitro is also a feature used to distinguish the two cell types. Cells of myeloid origin form in vitro colon
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