Experemental Study Of Brain Tumor Stem Cell Differentiationed In Vitro

Part one: The characteristics of morphology and differentiation related marker and proliferation dynamics of differentiated brain tumor stem cells in vitro[Objective]To pursue the change of cell morphology,expression of differentiation related markers ,and proliferation cycles of BTSC after differentiation in vitro.[Methods] Tumor stem cell spheres were from one clinical speciment(diagnosed anaplasia ependymocytoma).By magnetic sorting,CD133~+ cells were acquired. Cells differentiate in medium containing 10% fetal bovine serum(FBS).The morphology of cells were observed under phase contrast microscope.Cells were collected respectively before differentiation and in the 3rd,7th 10 th,21th day after differentiation.Cell surface markers such as CD133,Nestin,GFAP, β-TubulinIII were detected with flow cytometry.Proliferation cycles were examined before differentiation and in the 7th day after differentiation.Normal NSCs were taken as control.[Result] (1) cell morphology: The morphology of BTSC changed from round shape at the beginning,then turned to short fusiform, polygon and long fusiform.Seven days later,cells reversed to short fusiform and round shape.Cells accumulated into cell spheres and floated in culture medium again.While NSC differentiated along their routine rules.(2) Differentiation related markers:Both of undifferentiated BTSC and NSC showed high level expression of CD133 and Nestin. After differentiation,BTSC expressed CD133 and Nestin persistently,the expression level decreased first then increased. The expression ratio were (3.65±0.17) % and(28.99±1.26) % in the 7th day,(14.63±1.16)% and (45.46±1.27)% in the 21th day.While positive expression ratio of GFAP was higher than β-TubulinIII.The expression of CD133 and Nestin was lost when NSC were under differentiation in the 10 th day.The percentage of GFAP~+ cells and β-TubulinIII~+ cells were (88.94±1.23) %和 (11.94±0.36)% respectively. (3) Cell cycle and diploid: All undifferentiated BTSC were hypodiploid.After differentiation,majority of BTSC were hypodiploid or...