人脑胶质瘤干细胞扩增球体模型的建立及其蛋白质和脂质含量检测

Objective: The aim of this study is to establish a three-dimensional spheroid expansion model of human glioma stem cells with spontaneous spherogenesis and multidirectional differentiation potential in vitro, and to detect the contents of lipids, proteins and their secondary components by Raman spectroscopy, thus laying a foundation for further study of spheroid metabolism. Methods: The GSC23 and SU3 cells were cultured in serum-free stem cell culture medium, and then the cultures were fixed in polyformaldehyde solution. After dehydration, paraffin embedding and sectioning, HE staining, immunohistochemical staining with glioma-associated protein markers were performed, and also Raman spectroscopy was used to analyze the contentsof lipids, proteins and the minor components in spheroid tissues. Results: The formation of solid tumor-like three-dimensional tumor spheres was detected in the culture dishes of both stem cells.Immunohistochemistry results verified the expressions of all common marker proteins in glioblastoma multiforme. In addition, the results of Raman imaging showed that the contents of lipids, proteins and phenylalanine in all spheres were stable, and the difference was not statistically significant (p>0.05). Conclusion: In addition to the appearance characteristics and infinite expansion ability of three-dimensional solid tumors, glioma stem cell spheres cultured in vitro also have the ability to stably store essential energy for the metabolism of tumor cells such as fat and protein, which is expected to serve as an in vitro model to mimic the metabolic processes of glioma lipids and proteins in vivo.