Analysis on advantages of autofluorescence technique in screening malignant of glioma stem cells

Abstract

Objective To determine the efficiency of autofluorescence technique in screening of glioma stem cells (GSCs). Methods Since GSCs favor riboflavin, the agent was added to the culture medium of normal glioma stem cells. Then flow cytometry was employed to screen glioma stem cells with autofluorescence. Subsequently, the proliferation of the obtained cells was detected by CCK-8 assay. Finally, a mouse model of xenograft was established to observe the survive and tumor size and weight. The expression of Ki67, PCNA and Nestin in the tumor mass was detected by immunohistochemical assay. Results Glioma stem cells with autofluorescence were successfully screened out by flow cytometry. CCK-8 assay indicated that the proliferation of autofluorescence glioma stem cells was significantly stronger (P < 0.01). In vitro tumor-bearing experiments suggested that mice with xenograft of fluorescent stem cells had significantly reduced survival time and increased tumor volume and weight (P < 0.01). Immunohistochemical assay showed that the expression levels of Ki67, PCNA and Nestin in the tumor mass were significantly increased (P < 0.05). The proliferation and malignancy of the autofluorescence GSCs were stronger than glioma stem GL261 cells screened by CD133+ labeling. The CD133- GSCs with higher malignancy and invasion were also screened out with this method. Conclusion Autofluorescence technique can screen GSCs with high malignancy

Keywords

• glioma
• riboflavin
• glioma stem cells
• tumor stem cells