Activity of the human immortalized endothelial progenitor cell line HEPC-CB.1 supporting in vitro angiogenesis

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 HEPC-CB.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. Electronic supplementary material The online version of this article (10.1007/s11033-020-05662-6) contains supplementary material, which is available to authorized users.

Endothelial Cells as Tools to Model Tissue Microenvironment in Hypoxia-Dependent Pathologies

“…Both HEPC-CB.1 and HEPC-CB.2 were investigated by flow cytometry in order to assess their immunophenotype. It has been shown, that they are positive for the following markers: general stem cells (CD133), hematopoietic stem cells (CD13), non-hematopoietic stem cells (CD271), mesenchymal stem cells (CD90) and endothelial cell markers; mature as CD202b, VEGFR2, CD146 and stem-like or activated as CD105 [ 67 , 82 ]. Both cell lines are positive for CXCR4 (fusin, CD184); the unique receptor for the CXCL12 (SDF1 stromal cell-derived factor 1) chemokine, and weakly express markers of stem cells and endothelial cells: CD44 and CD15s.…”

“…However, it should be mentioned that there were differences between both cell lines. Firstly, the higher expression of CD133 and lower of CD271 was presented by HEPC-CB.2 as compared to HEPC-CB.1 [ 82 ]. IL-10 was secreted at a lower level by HEPC-CB.2 than second cell line [ 67 ].…”

From Primary MSC Culture of Adipose Tissue to Immortalized Cell Line Producing Cytokines for Potential Use in Regenerative Medicine Therapy or Immunotherapy

“…Angiogenesis was quantified by measurement of the mean mesh size, total length of capillary-like structures and number of nodes using ImageJ software. (NIH, Wayne Rasband, USA), as previously described [ 55 ].…”