Cardiology Stem Cell Center, The Heart Center, Rigshospitalet, University Hospital Copenhagen, Copenhagen, Denmark.
J Transl Med. 2013 Sep 18;11:219. doi: 10.1186/1479-5876-11-219.
Adipose-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.
ASCs stimulated with rhVEGF(A165) 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.
ASCs 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.
ASCs 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.
脂肪来源的基质细胞（ASCs）在血管内皮生长因子（VEGF）和血清饥饿的刺激下，应用于首例人体双盲安慰剂对照 MyStromalCell 试验中，作为治疗缺血性心脏病（IHD）的一种新的治疗选择。这项体外研究探讨了 VEGF 和血清饥饿对来自健康供体和 IHD 患者的 ASC 内皮分化能力的影响。
用 rhVEGF(A165) 在血清饥饿培养基中刺激 1 至 3 周的 ASCs 与血清饥饿（2%胎牛血清）或完全培养基（10%胎牛血清）中的 ASCs 进行比较。使用 qPCR、流式细胞术和免疫细胞化学法测量 VEGF 受体、内皮和干细胞标志物的表达。还测量了体外管形成和增殖。
与完全培养基中的 ASCs 相比，VEGF 刺激和血清饥饿培养基中的 ASCs 显著增加了转录因子 FOXF1、内皮标志物 vWF 和受体 VEGFR1 的转录。在所有条件下，ASCs 均保持干细胞特征。ASCs 在 VEGF 刺激和血清饥饿培养基中形成管。健康和患者 ASCs 之间唯一的区别是增殖率的变化。
IHD 患者和健康供体的 ASCs 通过血清饥饿同样倾向于向内皮方向分化，但 VEGF 刺激没有明显的附加效果。该治疗未导致完全的内皮分化，但向内皮谱系启动。
