Shah Q A, Tan X, Bi S, Liu X, Hu S
Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China.
Cell Prolif. 2014 Aug;47(4):290-8. doi: 10.1111/cpr.12111. Epub 2014 May 13.
This study was conducted to compare phenotypes and in vitro angiogenic capacity of putative endothelial progenitor cells (EPCs) derived from bone marrow (BM) and peripheral blood (PB), from an avian species.
Mononuclear cells were isolated from chicken BM and PB (BMMNCs and PBMNCs) and cultured in EGM-2 medium. Cells at days 7-14 were used for the experiments. Expression of progenitor and endothelial markers, number of Dil-ac-LDL/lectin dual-positive cells and adipogenic and osteogenic differentiation were determined. Migration and in vitro angiogenic ability between BMMNC- and PBMNC-derived cells were compared.
PBMNCs developed typical EPC appearance, with initial spindle shape followed by a cobblestone form, whereas BMMNC-derived cells retained their constitutive spindle-like morphology throughout the study. Cells derived from both sources expressed CD133, CD31 and VEGFR-2, although PBMNCs-derived cells had lower CD133 expression. Nevertheless, number of Dil-ac-LDL/lectin dual-positive cells did not differ between groups. Adipogenic and osteogenic lineages were verified in BMMNC- but not in PBMNC-derived cells. PBMNC-derived cells formed tubular networks on Matrigel. However, BMMNC-derived cells formed few tube-like structures, which were not morphologically comparable to those developed by their counterparts.
Our results suggest that so called EPCs derived from BMMNCs are not 'true' EPCs, supporting previous findings on mammals that BM may not serve as an optimal isolation source of EPCs.
本研究旨在比较源自禽类骨髓(BM)和外周血(PB)的假定内皮祖细胞(EPCs)的表型和体外血管生成能力。
从鸡的骨髓和外周血中分离出单核细胞(BMMNCs和PBMNCs),并在EGM-2培养基中培养。使用第7至14天的细胞进行实验。测定祖细胞和内皮标志物的表达、Dil-ac-LDL/凝集素双阳性细胞的数量以及成脂和成骨分化情况。比较BMMNCs和PBMNCs来源细胞之间的迁移和体外血管生成能力。
PBMNCs呈现出典型的EPC外观,最初为纺锤形,随后呈鹅卵石样形态,而BMMNCs来源的细胞在整个研究过程中保持其固有的纺锤样形态。两种来源的细胞均表达CD133、CD31和VEGFR-2,尽管PBMNCs来源的细胞CD133表达较低。然而,两组之间Dil-ac-LDL/凝集素双阳性细胞的数量没有差异。在BMMNCs来源的细胞中验证了成脂和成骨谱系,但在PBMNCs来源的细胞中未验证。PBMNCs来源的细胞在基质胶上形成管状网络。然而,BMMNCs来源的细胞形成的管状结构很少,在形态上与对应细胞形成的结构不可比。
我们的结果表明,源自BMMNCs的所谓EPCs不是“真正的”EPCs,这支持了之前关于哺乳动物的研究结果,即骨髓可能不是EPCs的最佳分离来源。