Sphere formation of adipose stem cell engineered by poly-2-hydroxyethyl methacrylate induces in vitro angiogenesis through fibroblast growth factor 2

A number of researchers have been reporting a wide range of in vitro and in vivo studies of cell engraftment to enhance angiogenesis using stem cells. Despite these efforts, studies involving three-dimensional (3D) culture method that mimics in vivo environment have not reached its peak yet. In this...

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Published in:Biochemical and biophysical research communications 2015-12, Vol.468 (1-2), p.372-379
Main Authors: Kim, Jong-Ho, Lim, I-Rang, Joo, Hyung Joon, Choi, Seung-Cheol, Choi, Ji-Hyun, Cui, Long-Hui, Im, Lisa, Hong, Soon Jun, Lim, Do-Sun
Format: Article
Language:English
Subjects:
Adipose stem cell
Adipose Tissue - cytology
Animals
Cell Movement
Cells, Cultured
Coated Materials, Biocompatible - metabolism
Fibroblast growth factor 2
Fibroblast Growth Factor 2 - metabolism
Humans
in vitro angiogenesis
Mice, Inbred C57BL
Neovascularization, Physiologic
Poly-2-hydroxyethyl methacrylate
Polyhydroxyethyl Methacrylate - metabolism
Sphere
Spheroids, Cellular - cytology
Spheroids, Cellular - metabolism
Stem Cells - cytology
Stem Cells - metabolism
Tissue Engineering
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Summary:A number of researchers have been reporting a wide range of in vitro and in vivo studies of cell engraftment to enhance angiogenesis using stem cells. Despite these efforts, studies involving three-dimensional (3D) culture method that mimics in vivo environment have not reached its peak yet. In this study, we investigated the change and effects on cellular angiogenic growth factors through sphere formation of adipose stem cell (ASC) which is engineered by poly-2-hydroxyethyl methacrylate (Poly-HEMA). First of all, we successfully induced sphere formation of ASC (sph-ASC) on Poly-HEMA coated plates. sph-ASC represented significantly higher expression levels of anti-apoptotic and hypoxic factors compared to monolayer adherent ASC (adh-ASC). Interestingly, sph-ASC showed higher mRNA levels of the following genes; CD31, CD144, vWF, IGF-2, MCP-1, PDGF-A, VEGF-A, VEGF-C, and FGF-2. In addition, mRNA expressions of angiogenic growth factor receptors such as Flk1, FGFR1, FGFR2, and Tie2 were elevated in sph-ASC. In protein level, Cytokine/Chemokines antibody array revealed a significant increase of FGF-2 in sph-ASC (3.17-fold) compared to adh-ASC. To investigate the effects of FGF-2 on sph-ASC, Matrigel angiogenic invasion assay showed significant reduced level of FGF-2 in FGF-2 siRNA transfected sph-ASC (2.27-fold) compared to negative control siRNA transfected sph-ASC. These findings suggest that Poly-HEMA coated plates induce sphere formation of ASC which has significantly higher expression of FGF-2, and plays a critical role as a major regulating growth factor of in vitro angiogenesis. •Sphere formation of ASC (sph-ASC) is successfully produced on Poly-HEMA coated plates.•Hypoxia induced by sph-ASC increases expression of angiogenic growth factors.•sph-ASC augments in vitro angiogenic potential via up-regulation of FGF-2.•FGF-2 is a major factor for in vitro angiogenic invasion of sph-ASC.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2015.10.083