Isolation and propagation of colon cancer stem cells in vitro

Background: Recent investigations in colon cancer biology suggest that cancer growth is driven by colon cancer stem cells (CSCs). CSCs are responsible for tumor initiation, maintenance, spreading and relapse. The aim of this study was to isolate and propagate tumorigenic colon CSCs in vitro. Methods: Tumor samples from colon cancers were subjected to mechanical and enzymatic disassociation. After passing of tumor cell suspensions through the cell strainers, tumor cells were counted and their viability was determined. Then tumor cells were plated in the special stem cells medium (contains serum-free DMEM/F12 supplemented by growth factors). Tumor cells growth was evaluated by inverted microscope. Findings: After about 3 weeks of plating in specific medium, colon CSCs were appeared as cell spheres whereas differentiated tumor cells and nonmalaginent cells were unable to grow. Colon cancer spheres were able to propagated and passaged in the specific stem cells medium. Conclusion: Isolation and propagation of tumorigenic colon CSCs in vitro would help to devise novel diagnostic and therapeutic methods. Isolated colon CSCs can be applied for the studying of cell signaling and assessment of the effect of anti-cancer drugs.