The characteristics of endothelial progenitor cells derived from mononuclear cells of rat bone marrow in different culture conditions
The characteristics of endothelial progenitor cells derived from mononuclear cells of rat bone marrow in different culture conditions
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不同培养条件下大鼠骨髓单个核细胞来源的内皮祖细胞的特性
DOI：
10.1007/s10616-010-9329-2
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发表时间：
2011-05-01
期刊：
影响因子：
2.2
通讯作者：
Qin, Shucun
中科院分区：
文献类型：
--
作者：
Yang, Nana;Li, Dawei;Qin, Shucun
骨髓来源的内皮祖细胞（EPCs）是一种异质性细胞。本研究旨在寻找诱导骨髓单个核细胞向内皮祖细胞分化的适宜条件。研究了大鼠骨髓单个核细胞在不同培养基、不同诱导时间和不同培养表面条件下的分化能力。研究了与分化的MNCs相关的细胞形态学和内皮生物标志物。我们的研究结果表明，在EGM-2 MV（内皮细胞基础培养基-2，加上SingleQuots的生长补充剂）中培养的MNCs形成了囊状形状，晚期EPC样形态，而在完全培养基（CM，M199，含10%FBS，20 ng/mL VEGF和10 ng/mL bFGF）中培养的MNCs显示了梭形形状，早期EPC样形态。两种形态的细胞都能够在体外掺入DiI-ac-LDL并结合凝集素。EGM-2 MV中培养的MNCs比CM中培养的MNCs表现出更高的增殖率和更高的eNOS表达。在EGM-2 MV中培养的MNCs具有在Matrigel上形成管的能力。流式细胞术结果表明，CD 133表达在第12天最高，并且在没有纤连蛋白包被的培养皿中培养的细胞中，FLK-1和CD 133阳性细胞数量在第20天出现最多。此外，不同形状的细胞之间CD 133、CD 31和FLK-1/CD 133的表达水平无明显差异。我们的实验表明，来自骨髓的MNCs可以在EGM-2 MV中分化为具有快速增殖能力的晚期EP样细胞。这些MNCs也可以在CM中分化为早期EP样细胞。此外，纤维连接蛋白可能不是EPCs在三代后分化为成熟EC所必需的。EGM-2 MV中骨髓来源的分化的MNCs具有EPC的特征，尽管EPC标志物的表达水平低于先前报道的水平。
Endothelial progenitor cells (EPCs) derived from bone marrow are known to be heterogeneous. In this study, we tried to find favorable conditions that induce the differentiation of mononuclear cells (MNCs) from bone marrow into EPCs. The differentiation capacity of MNCs from rat bone marrow was investigated in different conditions, such as different media, different induction times and different culture surfaces. The cell morphology and endothelial biomarkers associated with differentiated MNCs were studied. Our results indicated that MNCs cultured in EGM-2MV (Endothelial cell basal medium-2, plus SingleQuots of growth supplements) developed a bursiform shape, a late EPC-like morphology, while MNCs cultured in complete medium (CM, M199 with 10% FBS, 20 ng/mL VEGF and 10 ng/mL bFGF) showed a spindle shape, an early EPC-like morphology. Cells of both morphologies were able to incorporate DiI-ac-LDL and bind lectin in vitro. MNCs cultured in EGM-2MV exhibited a higher proliferation rate and higher eNOS expression than MNCs cultured in CM. MNCs cultured in EGM-2MV had the ability to form tubes on Matrigel. Flow cytometry results indicated that CD133 expression was highest at day 12 and that the greatest number of cells positive for both FLK-1 and CD133 appeared at day 20 from cells cultured in dishes without fibronectin coating. In addition, the expression levels of CD133, CD31 and FLK-1/CD133 were not significantly different between cells of different shapes. Our experiments suggest that MNCs from bone marrow can be differentiated into late EP-like cells in EGM-2MV, which have the ability to rapidly proliferate. These MNCs can also be differentiated into early EP-like cells in CM. Additionally, fibronectin may not be necessary for the differentiation of EPCs to mature ECs after three generations. Differentiated MNCs from bone marrow in EGM-2MV have the characteristics of EPCs, although the expression levels of EPC markers were lower than previously reported.