U251MG人胶质瘤细胞在器官型脑片共培养体系中侵袭的定量分析。

Quantitative analysis of U251MG human glioma cells invasion in organotypic brain slice co-cultures.

作者信息

Xu W-L, Wang Y, Wu J, Li G-Y

机构信息

Department of Neurosurgery, The First Affiliated Hospital of China Medical University, Shenyang, China.

出版信息

Eur Rev Med Pharmacol Sci. 2016 Jun;20(11):2221-9.

PMID:27338045

Abstract

OBJECTIVE

To develop an in vitro model under conditions that highly resemble the in vivo situation for searching new therapeutics targeting invasive glioma cells.

MATERIALS AND METHODS

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.

RESULTS

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.

CONCLUSIONS

The OBSC model is a reliable and easy-to-perform in vitro method to quantify the glioma invasion ability.

摘要

目的

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。

材料与方法

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。