Isolation Culture And Ultrastuctural Observation Of Tumor Stem Cells In Human Brain Glioma

OBJECTIVE To isolation and culture tumor stem cells in human glioma and to observe their biological characters and ultrastructures. METHODS Glioma tissue from neurosurgical operation were dissociated into single cells and cultured in serum free medium.Their proliferation and the growth course of cell spheres were observed. The tumor stem cells were induced into differentiate and compared with the cells in the tumor tissue.The expression of nestin was detected by immunofluorescence staining. Ultrastructural observation was performed by electron microscope. RES ULTS Brain tumor stem cell spheres expressing nestin were isolated and cultured from anaplastic astrocytoma and glioblastoma tissue with the ability to self-renew and proliferate. They could differenttiate into cells similar to that in the tumor tissue in serum medium. The spheres and microvillus on the surface were observed under scanning electron microscope. Under the transmission electron microscope, the conjunction among the cells within the neurospheres were tight, the ratio of nucleus to cytoplasm was very high, the cellular nucleus were multiple and the organelle in the cytoplasm was simple. CONCLUSIONS Tumor stem cells which can self-renew differentiate and express nestin are exist in human glioma. Their ultrastructures show the characters of immature and malignant cells. The present study can help to the further research on human brain glioma and tumor stem cells.