Assistance Publique-Hôpitaux de Paris, Hôpital Saint Louis, Unité de Thérapie Cellulaire, Université Paris Diderot, INSERM Unit UMRS940, Paris, France.
Cell Transplant. 2010;19(9):1143-55. doi: 10.3727/096368910X504487. Epub 2010 May 4.
Umbilical cord blood (CB) represents a main source of circulating endothelial progenitor cells (cEPCs). In view of their clinical use, in either the autologous or allogeneic setting, cEPCs should likely be expanded from CB kept frozen in CB banks. In this study, we compared the expansion, functional features, senescence pattern over culture, and in vivo angiogenic potential of cEPCs isolated from fresh or cryopreserved CB (cryoCB). cEPCs could be isolated in only 59% of cryoCB compared to 94% for fresh CB, while CB units were matched in terms of initial volume, nucleated and CD34(+) cell number. Moreover, the number of endothelial colony-forming cells was significantly decreased when using cryoCB. Once cEPCs culture was established, the proliferation, migration, tube formation, and acetylated-LDL uptake potentials were similar in both groups. In addition, cEPCs derived from cryoCB displayed the same senescence status and telomeres length as that of cEPCs derived from fresh CB. Karyotypic aberrations were found in cells obtained from both fresh and cryoCB. In vivo, in a hind limb ischemia murine model, cEPCs from fresh and cryoCB were equally efficient to induce neovascularization. Thus, cEPCs isolated from cryoCB exhibited similar properties to those of fresh CB in vitro and in vivo. However, the low frequency of cEPCs colony formation after cryopreservation shed light on the need for specific freezing conditions adapted to cEPCs in view of their future clinical use.
脐带血 (CB) 是循环内皮祖细胞 (cEPCs) 的主要来源。鉴于它们在自体或同种异体环境中的临床应用，cEPCs 应该可以从保存在 CB 库中的冷冻 CB 中扩增。在这项研究中，我们比较了从新鲜或冷冻 CB (cryoCB) 中分离的 cEPCs 的扩增、功能特征、衰老模式以及体内血管生成潜力。与新鲜 CB 相比，仅能从 59%的 cryoCB 中分离出 cEPCs，而 cryoCB 单位在初始体积、有核细胞和 CD34(+) 细胞数量方面是匹配的。此外，当使用 cryoCB 时，内皮集落形成细胞的数量显著减少。一旦建立了 cEPCs 培养，两组的增殖、迁移、管形成和乙酰化 LDL 摄取能力相似。此外，源自 cryoCB 的 cEPCs 显示出与源自新鲜 CB 的 cEPCs 相同的衰老状态和端粒长度。在新鲜和 cryoCB 获得的细胞中发现了染色体畸变。在体内，在后肢缺血小鼠模型中，来自新鲜和 cryoCB 的 cEPCs 同样有效地诱导血管新生。因此，源自 cryoCB 的 cEPCs 在体外和体内均表现出与新鲜 CB 相似的特性。然而，冷冻保存后 cEPCs 集落形成的低频率提示需要针对未来临床应用的特定冷冻条件来适应 cEPCs。
