Establishment of a new conditionally immortalized human skeletal muscle microvascular endothelial cell line
Establishment of a new conditionally immortalized human skeletal muscle microvascular endothelial cell line
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DOI：
10.1002/jcp.25772
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发表时间：
2017-12
影响因子：
5.6
通讯作者：
Hironori Sano;Y. Sano;Eri Ishiguchi;F. Shimizu;M. Omoto;T. Maeda;H. Nishihara;Y. Takeshita;Shiori Takahashi;Mariko Oishi;T. Kanda
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作者：
Hironori Sano;Y. Sano;Eri Ishiguchi;F. Shimizu;M. Omoto;T. Maeda;H. Nishihara;Y. Takeshita;Shiori Takahashi;Mariko Oishi;T. Kanda
在骨骼肌中，毛细血管具有紧密连接（TJs），其结构类似于血脑屏障（BBB）和血神经屏障（BNB）。虽然在BBB和BNB领域已经有了许多发现，但迄今为止对骨骼肌毛细血管tj的研究还很少。此外，目前还没有体外建立人体骨骼肌微血管模型的报道。我们新建立了一个新的人类骨骼肌微血管内皮细胞（HSMMEC）系。hsmmec从人类骨骼肌中分离出来，并被携带温度敏感的sv40t抗原和端粒酶基因的逆转录病毒感染。该细胞系被命名为TSM15，表现出纺锤状纤维形态，对抗因子VIII和抗VE钙粘蛋白抗体具有免疫反应性，并且对温度敏感。在33°C培养条件下，TSM15细胞稳定生长40代以上，从而保持了其纺锤状纤维形状的形态和融合时的接触抑制。这些细胞表达紧密连接的分子，如claudin‐5、occludin和occludens - 1，以及转运蛋白，如葡萄糖转运蛋白1。TSM15的跨内皮电阻与人脑微血管内皮细胞系相当。这种新的细胞系可能有助于炎性肌病（如皮肌炎）的病理生理分析，并可以提高我们对人体骨骼肌微血管生理生化特性的理解。
In skeletal muscle, the capillaries have tight junctions (TJs) that are structurally similar to those in the blood‐brain barrier (BBB) and blood‐nerve barrier (BNB). Although many findings have been clarified in the territory of BBB and BNB, few have so far examined the TJs of capillaries in the skeletal muscle. In addition, no in vitro human skeletal muscle microvasculature models have been reported thus far. We newly established a new human skeletal muscle microvascular endothelial cell (HSMMEC) line. HSMMECs were isolated from human skeletal muscle and were infected with retroviruses harboring temperature‐sensitive SV40 T antigen and telomerase genes. This cell line, termed TSM15, showed a spindle fiber‐shaped morphology, an immunoreactivity to anti‐factor VIII and anti‐VE‐cadherin antibodies, and a temperature‐sensitive growth. TSM15 cells grew stably for more than 40 passages when they were cultured at 33°C, thereby retaining their spindle fiber‐shaped morphology and contact inhibition at confluence. The cells expressed tight junctional molecules such as claudin‐5, occludin, and zonula occludens‐1, as well as transporters such as a glucose transporter 1. The transendothelial electrical resistance of TSM15 was as high as those of the human brain microvascular endothelial cell line. This novel cell line might facilitate the analyses of the pathophysiology of inflammatory myopathy, such as dermatomyositis, and can improve our understanding of the physiological and biochemical properties of the microvasculature in human skeletal muscle.