Isolation and identification of lung cancer stem cells from NCI-H1650 cell line / 重庆医学

ABSTRACT

Objective To isolate and identify the lung cancestem like cellfrom the cell line NCI-H1650 .MethodThe NCI-H1650 cellwere continuously cultured in serum-free medium with human insulin ,recombinanepidermal grow th facto(EGF) ,recombinanbasifibroblasgrowth facto(bFGF) and bovine serum albumin(BSA) ,and paclitaxel waadopted to induce the stem cell.Flow cytometry and immunofluorescenstaining were used to analyze the expression levelof lung cancestem cell markeCD133 and CD326 .The expressionof stem cell related markeOct4 ,Nanog and Sox-2 were detected by RT-Pcand West-ern blo.ResultThe NCI-H1650 cellcontinuously cultured by serum-free medium and induced by combining paclitaxel induction showed the spherical suspension growth .Compared with NCI-H1650 cellcultured by serum ,CD133 and CD326 had significantly high expression and the expression levelof Oct4 ,Nanog and Sox-2 were significantly increased (P＜0 .05) .Conclusion Adopting the serum-free culture in vitro combined with paclitaxel foinducing NCI-H1650 cellcan effectively isolate the stem cellamong them.