Three specific antigens to isolate endothelial progenitor cells from human liposuction material
Three specific antigens to isolate endothelial progenitor cells from human liposuction material
复制标题
DOI：
10.1016/j.jcyt.2013.06.018
复制
发表时间：
2013-11-01
期刊：
影响因子：
4.5
通讯作者：
Gabriel, Christian
中科院分区：
文献类型：
--
作者：
Hager, Gudrun;Holnthoner, Wolfgang;Gabriel, Christian
背景的目标。人内皮祖细胞（endothelial progenitor cells， EPC）在再生医学中发挥着重要作用，有助于血管损伤后的新生血管形成。它们通常从外周血、脐带血和骨髓中富集。在人类脂肪组织中，EPC是罕见的，其分离仍然是一个挑战。方法。采用胶原酶消化法制备脂肪组织，流式细胞术检测基质血管组分（SVF）中特异性标记蛋白的表达。富集时，用cd133微珠对细胞进行磁分选，EPC培养至菌落形成。用CD34进行第二次纯化；使用CD34和CD31微珠的组合进行额外的分离步骤。通过流式细胞术、Matrigel法和乙酰化低密度脂蛋白的摄取研究富集细胞内皮特异性标志物的表达。结果。表达模式证实了SVF的异质性，可以检测到罕见数量的cd133 +。经磁富集获得的内皮细胞EPC呈鹅卵石状，表达特异性标志物CD31、CD144、血管内皮生长因子（VEGF）R2、CD146、CD73和CD105。通过摄取乙酰化的低密度脂蛋白和在基质上形成管状结构，证实了功能的完整性。结论。通过磁性细胞分选，利用针对CD133（早期的EPC标记物）、CD34（干细胞标记物）和CD31（内皮细胞的典型标记物）的微珠组合，可以从人脂肪组织中富集稀有的EPC。在培养中，它们分化为EC，因此在再生医学中可能有促进新生血管形成的潜力。
Background aims. Human endothelial progenitor cells (EPC) play an important role in regenerative medicine and contribute to neovascularization on vessel injury. They are usually enriched from peripheral blood, cord blood and bone marrow. In human fat tissue, EPC are rare and their isolation remains a challenge. Methods. Fat tissue was prepared by collagenase digestion, and the expression of specific marker proteins was evaluated by flow cytometry in the stromal vascular fraction (SVF). For enrichment, magnetic cell sorting was performed with the use of CD 133 microbeads and EPC were cultured until colonies appeared. A second purification was performed with CD34; additional isolation steps were performed with the use of a combination of CD34 and CD31 microbeads. Enriched cells were investigated by flow cytometry for the expression of endothelial specific markers, by Matrigel assay and by the uptake of acetylated low-density lipoprotein. Results. The expression pattern confirmed the heterogeneous nature of the SVF, with rare numbers of CD 133+ detectable. EPC gained from the SVF by magnetic enrichment showed cobblestone morphology of outgrowth endothelial cells and expressed the specific markers CD31, CD144, vascular endothelial growth factor (VEGF)R2, CD146, CD73 and CD105. Functional integrity was confirmed by uptake of acetylated low-density lipoprotein and the formation of tube-like structures on Matrigel. Conclusions. Rare EPC can be enriched from human fat tissue by magnetic cell sorting with the use of a combination of microbeads directed against CD133, an early EPC marker, CD34, a stem cell marker, and CD31, a typical marker for endothelial cells. In culture, they differentiate into EC and hence could have the potential to contribute to neovascularization in regenerative medicine.