Centre for Nanotechnology and Regenerative Medicine, University College London Division of Surgery and Interventional Science, University College London, London, United Kingdom.
Biotechnol Appl Biochem. 2011 Sep-Oct;58(5):328-34. doi: 10.1002/bab.47. Epub 2011 Sep 23.
Human peripheral blood (HPB) contains both circulating endothelial cells (CECs) and endothelial progenitor stem cells (EPCs), which may be suitable for use in regenerative medicine. There has been considerable interest in using these cells, but there is no "gold standard" technique for isolating these cells. The aim of this study was to characterize and compare a number of different extraction and culture techniques to develop a system to isolate and culture cells. EPC and CEC were isolated from HPB using either Histopaque-1077 or Lymphoprep. The two isolation methods were compared for the number of cells isolated, cell metabolism, and RNA expression. Both isolations produced viable cells and were comparable. The tissue culture method employed does have a significant effect on the cell population with regard to medium choice, fetal bovine serum concentration, and surface modification of the culture surface. In conclusion, it can be seen that although this study and previous work can suggest a basis for culture, further work to develop an optimized and agreed "gold standard" culture regime for EPC from HPB is required to maximize the potential of this source of cells for regenerative medicine and to translate its clinical use in the future.
人外周血（HPB）中既含有循环内皮细胞（CECs），也含有内皮祖细胞（EPCs），这些细胞可能适用于再生医学。人们对使用这些细胞产生了浓厚的兴趣，但目前还没有用于分离这些细胞的“金标准”技术。本研究旨在对多种不同的提取和培养技术进行表征和比较，以开发一种分离和培养细胞的系统。使用 Histopaque-1077 或 Lymphoprep 从 HPB 中分离 EPC 和 CEC。比较了两种分离方法的细胞分离数量、细胞代谢和 RNA 表达。两种分离方法都能获得有活力的细胞，且效果相当。组织培养方法的选择对细胞群体有显著影响，包括培养基选择、胎牛血清浓度和培养表面的表面修饰。总之，虽然本研究和以前的工作可以为培养提供依据，但需要进一步研究以开发一种优化和公认的从 HPB 中分离 EPC 的“金标准”培养方案，以最大限度地发挥这种细胞来源在再生医学中的潜力，并为其未来的临床应用提供依据。
