Xu W-L, Wang Y, Wu J, Li G-Y
Department of Neurosurgery, The First Affiliated Hospital of China Medical University, Shenyang, China.
Eur Rev Med Pharmacol Sci. 2016 Jun;20(11):2221-9.
To develop an in vitro model under conditions that highly resemble the in vivo situation for searching new therapeutics targeting invasive glioma cells.
We generated organotypic brain slice "co-cultures" (OBSC) from mice and cultured the models on Millicell-CM membrane inserts. U251MG glioma cells expressing enhanced green fluorescent protein (EGFP) were established. After cultured the glioma cells to form spheroids, we implanted the spheroids onto brain slice surface. Then we evaluated the invasion area and cell density after U251MG cells were treated with the Na+-K+-2Cl- cotransporter 1 (NKCC1) inhibitor bumetanide by confocal laser microscopy.
In the models, the organotypic morphology and neuronal viability were well preserved. The confocal results showed that the cell spheroid area and density of U251MG cells in bumetanide group were decreased compared to the control group in brain slices. Meanwhile, the phospho-NKCC1(p-NKCC1) protein level of U251MG cells in bumetanide-treated group was also lower than the control group.
The OBSC model is a reliable and easy-to-perform in vitro method to quantify the glioma invasion ability.
构建一种高度类似于体内情况的体外模型，以寻找针对侵袭性胶质瘤细胞的新疗法。
我们从小鼠中制备了器官型脑片“共培养物”（OBSC），并在Millicell-CM膜插入物上培养这些模型。建立了表达增强型绿色荧光蛋白（EGFP）的U251MG胶质瘤细胞。在将胶质瘤细胞培养形成球体后，我们将球体植入脑片表面。然后，通过共聚焦激光显微镜评估用Na+-K+-2Cl-共转运体1（NKCC1）抑制剂布美他尼处理U251MG细胞后的侵袭面积和细胞密度。
在这些模型中，器官型形态和神经元活力得到了很好的保留。共聚焦结果显示，与脑片中的对照组相比，布美他尼组中U251MG细胞的细胞球体面积和密度降低。同时，布美他尼处理组中U251MG细胞的磷酸化NKCC1（p-NKCC1）蛋白水平也低于对照组。
OBSC模型是一种可靠且易于操作的体外方法，可用于量化胶质瘤的侵袭能力。
