Enrichment of breast cancer stem cells using a keratinocyte serum-free medium
10.3760/cma.j.issn.0366-6999.2011.18.025
- Author:
Zhen-Zhen LIU
1
,
2
;
Ping CHEN
;
Zhen-Duo LU
;
Shu-De CUI
;
Zi-Ming DONG
Author Information
1. Department of Pathophysiology, Basic Medical College,Zhengzhou University, Zhengzhou, Henan 450053, China
2. Breast Cancer Center, Henan Cancer Hospital and Affiliated Cancer Hospital of Zhengzhou University, Zhengzhou, Henan450008, China
- Keywords:
breast cancer;
cancer stem cells;
keratinocyte serum-free medium;
defined medium,enrichment
- From:
Chinese Medical Journal
2011;124(18):3934-2936
- CountryChina
- Language:Chinese
-
Abstract:
Background Keratinocyte serum-free medium (K-SFM) is a defined medium used to support the growth of primary keratinocytes and embryonic stem cell. The aim of this research was to optimize enrichment of breast cancer stem cells (CSCs) using K-SFM.Methods A K-SFM was used to enrich CSCs from two breast cancer cell lines and a primary culture of breast cancer.RPMI-1640 supplemented with 10％ fetal calf serum (FCS) was used as a control. CSCs were identified with flow cytometry using CD44+/CD24-as molecular markers. The expression of a variety of CSC markers (Oct-4, ABCG2, Nanog,N-cadherin, and E-cadherin) was analyzed with real-time PCR.Results Much higher percentage of CSCs was achieved with K-SFM: 17.3％ for MCF-7 cells, 17.4％ for SKBR-3, and 20.0％ for primary breast cancer culture. Less than 1％ CSC was achieved using RPMI-1640 supplemented with 10％ FCS. In comparison to the CSCs obtained with RPMI-1640, CSCs in the K-SFM expressed higher levels of Oct-4,ABCG2, Nanog and N-cadherin, and lower level of E-cadherin.Conclusion K-SFM is an optimal culture medium to maintain and to enrich breast CSCs.
