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A modified method by differential adhesion and serum-free culture medium for enrichment of cancer stem cells


1 Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Nanfang Hospital; Department of Urology, Nanfang Hospital, Southern Medical University, Guangzhou, China
2 Department of Neurology, TCM-Integrated Cancer Center of Southern Medical University, Guangzhou, China
3 Department of Urology, Nanfang Hospital, Southern Medical University, Guangzhou, China

Correspondence Address:
Wan-Long Tan,
Department of Urology, Nanfang Hospital, Southern Medical University, Guangzhou
China
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Source of Support: None, Conflict of Interest: None

Objective: In this study, we showed a modified method for the isolation of cancer stem cells (CSCs) using a combination of differential adhesion method and serum-free culture medium (SFM) method. Materials and Methods: Trypsin-sensitive cells and trypsin-resistant cells were isolated from MB49, EJ, and SK-OV-3 cells using a combination of differential adhesion method and SFM method. The CSCs markers expression of trypsin-resistant cells was verified by the flow cytometry, the Western blotting, and the quantitative polymerase chain reaction. Functional comparisons were verified by the resistance to chemotherapy assay, the transwell assay, and the tumor xenograft formation assay. Results: Trypsin-resistant cells were isolated successfully. They were identified with high expression of CSCs markers and possessed higher resistance to chemotherapy, greater migration in vitro and stronger tumorigenic abilities in vivo. Conclusion: Trypsin-resistant cells showed specific CSCs characterizations. They were able to be isolated successfully with a modified method by a combination of differential adhesion method and SFM method.


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    -  Zhu YT
    -  Wang CY
    -  Pang SY
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