Investigation of index of therapeutic modes as cell drugs in human mesenchymal stem cells and their validation

我们研究了可作为预测缺血条件下人骨髓间充质干细胞(HMSCs)分泌血管内皮生长因子(VEGF)水平的候选基因。在正常和缺血条件下培养人骨髓间充质干细胞。24小时后收集细胞上清液，用酶联免疫吸附试验检测血管内皮细胞生长因子。我们分析了6批hMSCs缺血前基因表达(Affymetrix HG-U133Plus2.0)与缺血后血管内皮生长因子分泌的相关性。17个基因在缺血前基因表达与缺血后VEGF水平(量或率的变化)的组合中均呈正相关。此外，针对17个基因中的2个的RNAi减少了对缺血的血管内皮生长因子的分泌。在本研究中，17个基因被确定为评估缺血后hMSCs质量和疗效的候选基因。另外，有2个基因的表达下调降低了血管内皮细胞生长因子的反应，从而促进了血管内皮细胞在缺血状态下的分泌。

We investigated the candidate genes which can become a marker to predict vascular endothelium growth factor (VEGF) secretional level of human mesenchymal stem cells (hMSCs)under ischemia. hMSCs were cultured under normal condition or ischemic condition. Cell supernatant was collected after 24hr and VEGF were measured with ELISA. We analyzed the correlation between gene expression before ischemia (Affymetrix HG-U133Plus2.0) and VEGF secretion after ischemia in hMSCs among six lots. 17 genes showed positive correlation in all combination of pre-ischemic gene expression and post-ischemic VEGF level (the change of amount or rate). Furthermore, RNAi against 2 of the 17 genes reduced VEGF secretion in response to ischemia. In this study, 17 genes were identified as candidate genes which could assess quality and efficacy of hMSCs after ischemia. In addition, it is suggested that 2 genes, the knockdown of which decrease the VEGF response, contribute to enhance VEGF secretion under ischemia.