Construction and characterization of osteogenic and vascular endothelial cell sheets from rat adipose-derived mesenchymal stem cells
Construction and characterization of osteogenic and vascular endothelial cell sheets from rat adipose-derived mesenchymal stem cells
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大鼠脂肪间充质干细胞成骨和血管内皮细胞片的构建和表征
DOI：
10.1016/j.tice.2016.07.004
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发表时间：
2016-10-01
期刊：
影响因子：
2.6
通讯作者：
An, Yilin
中科院分区：
文献类型：
--
作者：
Zhang, Hualin;Yu, Na;An, Yilin
本研究从大鼠脂肪组织中分离脂肪源性间充质干细胞（ADSCs）。流式细胞术鉴定显示，3代ADSCs高表达CD29和CD44，但几乎不表达CD31和CD45。分别通过油红0染色、碱性磷酸酶（ALP）和阿利新蓝染色证实成脂、成骨和软骨分化。在成骨培养基中培养密度为1 × 10(6)/cm(2)的ADSCs， 14d后可获得成骨细胞片，von kossa染色阳性。透射电镜（TEM）结果表明，ECM表面沉积有针状钙盐晶体。这些结果提示成骨细胞片可能具有潜在的成骨能力。在内皮细胞生长培养基中培养密度为1 × 10(6)/cm(2)的ADSCs，培养16 d后可形成内皮细胞片。透射电镜图像证实细胞内可见Weibel-Palade小体。CD31表达阳性，提示内皮细胞片可能具有较强的血管形成能力。本研究通过体外诱导培养ADSCs获得了两种具有成骨和血管形成潜能的细胞片，为构建血管化组织工程骨治疗骨缺损奠定了基础。(C) 2016 Elsevier Ltd.版权所有。
In this study, adipose-derived mesenchymal stem cells (ADSCs) were isolated from adipose tissues of rats. Flow cytometry identification showed that ADSCs of passage 3 highly expressed CD29 and CD44, but hardly expressed CD31 and CD45. Adipogenic, osteogenic, and chondrogenic differentiation were confirmed by the results of oil red 0 staining, alkaline phosphatase (ALP), and alcian blue staining, respectively. ADSCs at a density of 1 x 10(6)/cm(2) were cultured in the osteogenic medium and the osteogenic cell sheets could be obtained after 14 d. The cell sheets were positive with von kossa staining. The transmission electron microscopy (TEM) result showed that needle-like calcium salt crystals were deposited on the ECM. These results suggested that the osteogenic cell sheets may have potential osteogenesis ability. ADSCs at a density of 1 x 10(6)/cm(2) were cultured in the endothelial cell growth medium-2 and the endothelial cell sheets can be formed after 16 d of culture. The TEM image confirmed that the Weibel-Palade corpuscle was seen in the cells. The expression of CD31 was positive, suggesting that the endothelial cell sheets may have a strong ability to form blood vessels. In this study, two types of cell sheets with the potential abilities of osteogenesis and blood vessels formation were obtained by induced culture of ADSCs in vitro, which lays a foundation to build vascularized tissue engineered bone for the therapy of bone defects. (C) 2016 Elsevier Ltd. All rights reserved.