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

本研究旨在寻找能够预测缺血状态下人骨髓间充质干细胞（hMSCs）血管内皮生长因子（VEGF）分泌水平的候选基因。在正常条件和缺血条件下培养hMSCs。24小时后收集细胞上清液，用ELISA测定VEGF。分析6批hMSCs缺血前基因表达（AffyphineHG-U133 Plus 2.0）与缺血后VEGF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）的所有组合中均呈正相关。此外，针对17个基因中的2个的RNAi减少了响应于缺血的VEGF分泌。在本研究中，17个基因被确定为候选基因，可以评估缺血后的hMSCs的质量和疗效。此外，它表明，2个基因，敲低降低VEGF的反应，有助于增强VEGF分泌缺血。

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.