In previous studies, vascular smooth muscle progenitor cells (vSMPCs) isolated from peripheral blood mononuclear cells (PBMCs) were cultured using medium containing platelet-derived growth factor-BB (PDGF-BB) for 4 weeks. However, this method requires long culture periods of up to 4 weeks and yields low cell counts. Therefore, we proposed the modified method to improve the cell yield and purity and to reduce the cell culture period. PBMCs were isolated from human peripheral blood and cultured by the conventional method using medium containing PDGF-BB alone or the modified method using medium containing PDGF-BB, basic fibroblast growth factor (bFGF), and insulin-transferrin-selenium ITS for 4 weeks. The purity of vSMPCs was analyzed for the expression of a- smooth muscle actin (SMA) by flow cytometry and significantly higher in the modified method than conventional methods at the 1st and 2nd weeks. Also, mRNA expression of a-SMA by real-time PCR was significantly higher in the modified method than conventional method at the 2 weeks. The yield of vSMPCs by trypan blue exclusion assay was significantly higher in the modified method than conventional method at the 1st, 2nd and 3rd weeks. The primary culture using the modified method with PDGF-BB, bFGF, and ITS not only improved cell purity and yield, but also shortened the culture period, compared to the conventional culture method for vSMPCs. The modified method will be a time-saving and useful tool in various studies related to vascular pathology.

在以前的研究中，使用含有血小板衍生生长因子-BB（PDGF-BB）的培养基培养从外周血单核细胞（PBMC）分离出的血管平滑肌祖细胞（vSMPC）4周。但是，这种方法需要长达4周的长时间培养，并且细胞计数低。因此，我们提出了改进的方法以提高细胞产量和纯度并减少细胞培养时间。从人外周血中分离PBMC，并通过常规方法使用单独包含PDGF-BB的培养基进行培养，或使用包含PDGF-BB，碱性成纤维细胞生长因子（bFGF）和胰岛素-转铁蛋白-硒ITS的培养基进行改良方法培养4周。通过流式细胞术分析了vSMPC的纯度，以检测a-平滑肌肌动蛋白（SMA）的表达，在改良方法中，第1周和第2周的水平明显高于传统方法。此外，在2周内，改良方法比常规方法的实时PCR检测a-SMA的mRNA表达明显更高。在第1、2、3周，改良方法中锥虫蓝排除法测定的vSMPC的产量明显高于传统方法。与vSMPC的常规培养方法相比，使用带有PDGF-BB，bFGF和ITS的改良方法进行的原代培养不仅提高了细胞纯度和产量，而且缩短了培养周期。改进后的方法将在各种与血管病理学相关的研究中节省时间并成为有用的工具。