Mechanotransduction Effects on Endothelial Cell Proliferation via CD31 and VEGFR2: Implications for Immunomagnetic Separation.
Kalpesh D MahajanGauri M NabarWei XueMirela AnghelinaNicanor I MoldovanJeffrey J ChalmersJessica O WinterPublished in: Biotechnology journal (2017)
Immunomagnetic separation is used to isolate circulating endothelial cells (ECs) and endothelial progenitor cells (EPCs) for diagnostics and tissue engineering. However, potentially detrimental changes in cell properties have been observed post-separation. Here, the effect of mechanical force, which is naturally applied during immunomagnetic separation, on proliferation of human umbilical vein endothelial cells (HUVEC), kinase insert domain-positive receptor (KDR) cells, and peripheral blood mononuclear cells (PBMCs). Cells are exposed to CD31 or Vascular Endothelial Growth Factor Receptor-2 (VEGFR2) targeted MACSi beads at varying bead to cell ratios and compared to free antibody and unconjugated beads. A vertical magnetic gradient is applied to static 2D cultures, and a magnetic cell sorter is used to analyze cells in dynamic flow. No significant difference in EC proliferation is observed for controls or VEGFR2-targeting beads, whereas CD31-conjugated beads increase proliferation in a dose dependent manner in static 2-D cultures. This effect occurs in the absence of magnetic field, but is more pronounced with magnetic force. After flow sorting, similar increases in proliferation are seen for CD31 targeting beads. Thus, the effects of targeting antibody and magnetic force applied should be considered when designing immunomagnetic separation protocols for ECs.
Keyphrases
- vascular endothelial growth factor
- endothelial cells
- induced apoptosis
- signaling pathway
- cell cycle arrest
- liquid chromatography
- single cell
- cell proliferation
- molecularly imprinted
- cancer therapy
- cell therapy
- endoplasmic reticulum stress
- single molecule
- high glucose
- tissue engineering
- cell death
- mass spectrometry
- drug delivery
- oxidative stress
- stem cells
- cell cycle
- tandem mass spectrometry
- high resolution
- simultaneous determination
- bone marrow