Experimental study of real-time monitoring of glioma stem cell differentiation process

Abstract

Objective To observe the changes in a variety of molecular markers in the differentiation process from glioma stem cells to endothelial cells using live cell fluorescence imaging technique. Methods Live cell fluorescence imaging technique was used to study the differentiation process of glioma stem cell GSC-1 for dynamic observation of 12 h. The immunofluorescence staining was used to perform endothelial cell surface marker CD34 staining during the three dimensional culture at 0, 2, and 6 h, and the morphological changes of GSC-1 were observed by light microscopy. Results Under the three-dimensional culture, GSC-1 cells could form tube-like structures. The number of some cells expressed endothelial cell markers CD34, CD34 positive cells increased with the time of differentiation (P<0.05), while stem marker disappeared gradually. Live cell fluorescence imaging observed that the number of CD34 positive cells increased with the time of induction and differentiation under the same field of vision. The whole process of a single GSC-1 cell transformed from a CD34 negative cell to a CD34 positive cell was observed. Conclusions The live cell fluorescence imaging technique detecting the cell differentiation process can get the same result of the traditional immunofluorescence assay, and it is more intuitive and accurate. It can follow up and observe a single cell for a long time. Key words: Glioma; Glioblastoma; Neoplastic stem cells; Cell differentiation