Identical effects of VEGF and serum-deprivation on phenotype and function of adipose-derived stromal cells from healthy donors and patients with ischemic heart disease

Abstract

BackgroundAdipose-derived stromal cells (ASCs) stimulated with vascular endothelial growth factor (VEGF) and serum-deprived, are applied in the first in-man double-blind placebo-controlled MyStromalCell Trial, as a novel therapeutic option for treatment of ischemic heart disease (IHD). This in vitro study explored the effect of VEGF and serum deprivation on endothelial differentiation capacity of ASCs from healthy donors and IHD patients.MethodsASCs stimulated with rhVEGFA165 in serum-deprived medium for one to three weeks were compared with ASCs in serum-deprived (2% fetal bovine serum) or complete medium (10% fetal bovine serum). Expression of VEGF receptors, endothelial and stem cell markers was measured using qPCR, flow cytometry and immunocytochemistry. In vitro tube formation and proliferation was also measured.ResultsASCs from VEGF-stimulated and serum-deprived medium significantly increased transcription of transcription factor FOXF1, endothelial marker vWF and receptor VEGFR1 compared with ASCs from complete medium. ASCs maintained stem cell characteristics in all conditions. Tube formation of ASCs occurred in VEGF-stimulated and serum-deprived medium. The only difference between healthy and patient ASCs was a variation in proliferation rate.ConclusionsASCs from IHD patients and healthy donors proved equally inclined to differentiate in endothelial direction by serum-deprivation, however with no visible additive effect of VEGF stimulation. The treatment did not result in complete endothelial differentiation, but priming towards endothelial lineage.

Human adipose-derived stromal cells in a clinically applicable injectable alginate hydrogel: Phenotypic and immunomodulatory evaluation

“…Isolation and culture expansion of ASCs isolated form abdominal adipose tissue was performed as described earlier [23] The remaining cells were centrifuged at 1200g for 10 min at room temperature, re-suspended and counted using NucleoCounter NC-100 (Chemometec) according to the manufacturer's instructions.…”

Hypoxic and ischemic effects on gene and protein expression levels of paracrine factors by human olfactory mucosa mesenchymal-like stem cells

“…23 Serum deprivation changes the expression of endothelial markers and the secretion of paracrine factors in MSCs. 24,25 As a population of the MSCs, olfactory mucosa mesenchymal-like stem cells (OM-MSCs) may work effectively in the treatment of cerebral ischemia. Olfactory mucosa is a self-renewal tissue, containing olfactory ensheathing cells (OECs) and OM-MSCs.…”

Identification of a common reference gene pair for qPCR in human mesenchymal stromal cells from different tissue sources treated with VEGF

“…vWF was chosen as positive control since it has been shown to be upregulated in other experiments by the same treatment, namely serum-deprivation combined with VEGF stimulation [19,22]. vWF was normalized to RGs from each selection step of our procedure.…”