Kwon Hyuk Min, Hur Sung-Mo, Park Keon-Young, Kim Chun-Ki, Kim Yong-Man, Kim Hyun-Soo, Shin Ha-Cheol, Won Moo-Ho, Ha Kwon-Soo, Kwon Young-Guen, Lee Dong Heon, Kim Young-Myeong
Department of Obstetrics and Gynecology, Kangwon National University Hospital, Chuncheon, Gangwon-do 200-701, Republic of Korea.
Departments of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do 200-701, Republic of Korea.
Vascul Pharmacol. 2014 Oct;63(1):19-28. doi: 10.1016/j.vph.2014.06.004. Epub 2014 Jul 2.
The therapeutic effects of stem cell transplantation in ischemic disease are mediated by the production of paracrine bioactive factors. However, the bioactive factors secreted by human mesenchymal stem cells (hMSCs) and their angiogenic activity are not clearly identified or determined. We here found that hMSC-derived conditioned media (hMSC-CdM) stimulated in vitro angiogenic activity of endothelial cells and contained significant levels of various growth factors and cytokines, such as vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), monocyte chemotactic protein-1 (MCP-1), interleukin-6 (IL-6), and transforming growth factor-beta1 (TGF-β1). The angiogenic activity of hMSC-CdM was significantly inhibited by pretreatment with neutralizing antibodies against VEGF, MCP-1, and IL-6, but not against TGF-β1 and HGF. A mixture of those inhibitory antibodies blocked CdM-mediated activation of angiogenic signals, as well as inhibited CdM-mediated in vivo angiogenesis. Moreover, local injection of CdM increased angiogenesis and promoted blood flow in mice with hindlimb ischemia, and these effects were inhibited by co-treatment with these inhibitory antibodies. These results indicate that hMSC-CdM represents a promising cell-free therapeutic strategy for neovascularization in ischemic diseases. These results suggest the combination of VEGF, MCP-1, and IL-6 as a commercial application for therapeutic angiogenesis.
干细胞移植在缺血性疾病中的治疗作用是由旁分泌生物活性因子介导的。然而，人骨髓间充质干细胞（hMSCs）分泌的生物活性因子及其血管生成活性尚未得到明确鉴定或确定。我们在此发现，hMSC条件培养基（hMSC-CdM）可刺激内皮细胞的体外血管生成活性，并含有大量的各种生长因子和细胞因子，如血管内皮生长因子（VEGF）、肝细胞生长因子（HGF）、单核细胞趋化蛋白-1（MCP-1）、白细胞介素-6（IL-6）和转化生长因子-β1（TGF-β1）。用抗VEGF、MCP-1和IL-6的中和抗体预处理可显著抑制hMSC-CdM的血管生成活性，但对TGF-β1和HGF无抑制作用。这些抑制性抗体的混合物可阻断CdM介导的血管生成信号激活，并抑制CdM介导的体内血管生成。此外，局部注射CdM可增加后肢缺血小鼠的血管生成并促进血流，而这些作用可被这些抑制性抗体联合处理所抑制。这些结果表明，hMSC-CdM是一种有前途的用于缺血性疾病新生血管形成的无细胞治疗策略。这些结果提示VEGF、MCP-1和IL-6的组合可作为治疗性血管生成的商业应用。
