背景:巨噬细胞增多是一种用于细胞外液内化的途径,白蛋白和溶解的分子。过去,由于手动采集和视觉评估图像的结合非常费力,因此评估巨细胞增多症一直具有挑战性。使得这种类型的评估对于高通量应用来说很困难。因此,有必要开发敏感和特异性的巨噬细胞增多症评估方法.
方法:本文提出了一种基于高含量分析(HCA)的定量,省时的巨噬细胞增多症检测方法。此外,使用CCK8试验评估细胞增殖。
结果:定义术语“巨藻细胞指数”以估计巨藻细胞增多并允许不同细胞系和治疗之间的比较。此外,我们证明,在类似肿瘤微环境的L-谷氨酰胺(L-Gln)缺乏的条件下,巨细胞增殖可以促进胶质母细胞瘤(GBM)细胞的存活.
结论:HCA代表了一种新的,非主观性和高通量试验用于巨噬细胞增多症评估。此外,L-Gln剥夺增加了GBM细胞中的宏染色体指数,这表明该过程可用于设计GBM疗法。
■支持本文结论的数据集包含在文章及其补充材料中。
Macropinocytosis is a pathway utilized for the internalization of extracellular fluid, albumin and dissolved molecules. Assessing macropinocytosis has been challenging in the past because the combination of manual acquisition and visual evaluation of images is laborious, making this type of assessment difficult for high-throughput applications. Therefore, there is a need to develop sensitive and specific macropinocytosis evaluation methods.
This paper proposed a quantitative and time-saving method for macropinocytosis detection based on high-content analysis (HCA). Additionally, cell proliferation was evaluated using CCK8 test.
The term \"macropinosome index\" was defined to estimate macropinocytosis and allow comparisons between different cell lines and treatments. Furthermore, we demonstrated that macropinocytosis can promote glioblastoma (GBM) cell survival under L-glutamine (L-Gln)-deficient conditions that resemble the tumour microenvironment.
HCA represents a novel, nonsubjective and high-throughput assay for macropinocytosis assessment. In addition, L-Gln deprivation increased the macropinosome index in GBM cells, suggesting that this process may be used to design GBM therapies.
The datasets supporting the conclusions of this article are included within the article and its supplementary materials.