The effect of glioma stem/progenitor cell on tumor angiogenesis traced by red/green fluorescent protein

Abstract

Objective To explore the potential role of glioma stem cell in the generation and maintenance of tumor blood vessels.Methods Gliomas stem cells (CD133 + tumor cells) transfected with red fluorescent protein (RFP) gene were implanted in situ stereotaxicly (25 μl,1 × 106 cells) or subcutaneously (200 μ1,1 × 106 cells) in nude mice,then the xenografts (2-3 weeks)were pressed fiat and tumor vessels structures were observed under fluorescence microscope and laser confocal microscope.Results Tissue squash method can clearly display the host normal vessels and some cells source of vascular endothelial cells inside the tumor region on the basis of different colors under fluorescence、laser confocal microscope.Images displayed vascular cells can be classified colors into three types:host derived cells(green),tumor derived(red),tumor derived and host-derived fusion(yellow).Cells form of tumor angiogenesis can be divided into three types:proliferation of endothelial cells(EC),tumor stem cells (TC),tumor stem cells and host-derived endothelial cells fusion (EC/TC).Conclusion Gliomas stem cells transfected with red fluorescent protein (RFP) gene were implanted in green fluorescent protein(GFP) transgenic nude mice model was very useful to study the processes of malignant glioma angiogenesis.This model can prove the glioma stem/progenitor cells act as initiating cell in tumor angiogenesis. Key words: Tumor stem cells; Fluorescent protein gene transfection; Transgenic fluorescence nude mice; Transplantable tumor; Tumor angiogenesis