Detection of CD133 (prominin-1) in a human hepatoblastoma cell line (HuH-6 clone 5).
Detection of CD133 (prominin-1) in a human hepatoblastoma cell line (HuH-6 clone 5).
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DOI：
10.1002/jemt.22237
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发表时间：
2013-08
影响因子：
2.5
通讯作者：
Nishiyama, Hidetoshi
中科院分区：
文献类型：
--
作者：
Akita, Masumi;Tanaka, Kayoko;Murai, Noriko;Matsumoto, Sachiko;Fujita, Keiko;Takaki, Takashi;Nishiyama, Hidetoshi
我们检查了人肝母细胞瘤细胞系（HuH-6 克隆 5）中的 CD133 分布。我们使用新开发的大气扫描电子显微镜（ASEM）直接观察缓冲溶液中耐压薄膜（氮化硅薄膜）上培养的细胞，该显微镜具有一个开放的样品皿，底部有氮化硅薄膜窗口，扫描电子显微镜束通过该窗口从下方扫描溶液中的样品。 ASEM 能够观察腹侧细胞表面，这是使用标准 SEM 无法观察到的。然而，用 ASEM 观察背侧细胞表面很困难。因此，我们开发了一种使用 Aclar® 塑料薄膜观察细胞背面的新方法。在此方法中，细胞在 Aclar 塑料膜上培养，细胞的背面与 ASEM 培养皿的氮化硅薄膜接触。使用 ASEM 进行的初步研究表明 CD133 主要位于细胞外围区域的膜皱褶中。标准透射电子显微镜和扫描电子显微镜显示CD133优先集中在包含丝状伪足和片状伪足前缘的复杂结构中。我们还观察到 CD133 与 F-肌动蛋白的共定位。抗 CD133 的抗体减少了细胞迁移。甲基-β-环糊精处理降低了细胞粘附以及片状足和丝状足的形成。胆固醇水平的降低可能会扰乱 CD133 膜定位。结果表明CD133膜定位在肿瘤细胞粘附和迁移中发挥作用。
We examined CD133 distribution in a human hepatoblastoma cell line (HuH-6 clone 5). We directly observed the cultured cells on a pressure-resistant thin film (silicon nitride thin film) in a buffer solution by using the newly developed atmospheric scanning electron microscope (ASEM), which features an open sample dish with a silicon nitride thin film window at its base, through which the scanning electron microscope beam scans samples in solution, from below. The ASEM enabled observation of the ventral cell surface, which could not be observed using standard SEM. However, observation of the dorsal cell surface was difficult with the ASEM. Therefore, we developed a new method to observe the dorsal side of cells by using Aclar® plastic film. In this method, cells are cultured on Aclar plastic film and the dorsal side of cells is in contact with the thin silicon nitride film of the ASEM dish. A preliminary study using the ASEM showed that CD133 was mainly localized in membrane ruffles in the peripheral regions of the cell. Standard transmission electron microscopy and scanning electron microscopy revealed that CD133 was preferentially concentrated in a complex structure comprising filopodia and the leading edge of lamellipodia. We also observed co-localization of CD133 with F-actin. An antibody against CD133 decreased cell migration. Methyl-β-cyclodextrin treatment decreased cell adhesion as well as lamellipodium and filopodium formation. A decrease in the cholesterol level may perturb CD133 membrane localization. The results suggest that CD133 membrane localization plays a role in tumor cell adhesion and migration.
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影响因子：
8.4
作者：
Monzani, Elena;Facchetti, Floriana;La Porta, Caterina A. M.
通讯作者：
La Porta, Caterina A. M.
影响因子：
64.8
作者：
O'Brien, Catherine A.;Pollett, Aaron;Dick, John E.
通讯作者：
Dick, John E.
影响因子：
3
作者：
Nishiyama, Hidetoshi;Suga, Mitsuo;Sato, Chikara
通讯作者：
Sato, Chikara
影响因子：
2.2
作者：
Suga, Mitsuo;Nishiyama, Hidetoshi;Sato, Chikara
通讯作者：
Sato, Chikara
影响因子：
21.3
作者：
Röper, K;Corbeil, D;Huttner, WB
通讯作者：
Huttner, WB