Enrichment of breast cancer stem cells using a keratinocyte serum-free medium.
- Author:
Zhen-Zhen LIU
1
;
Ping CHEN
;
Zhen-Duo LU
;
Shu-de CUI
;
Zi-Ming DONG
Author Information
- Publication Type:Journal Article
- MeSH: ATP Binding Cassette Transporter, Sub-Family G, Member 2; ATP-Binding Cassette Transporters; genetics; Cadherins; genetics; Cell Culture Techniques; methods; Cell Line, Tumor; Culture Media, Serum-Free; Female; Homeodomain Proteins; genetics; Humans; Keratinocytes; cytology; Nanog Homeobox Protein; Neoplasm Proteins; genetics; Neoplastic Stem Cells; cytology; metabolism; Octamer Transcription Factor-3; genetics; Real-Time Polymerase Chain Reaction
- From: Chinese Medical Journal 2011;124(18):2934-2936
- CountryChina
- Language:English
-
Abstract:
BACKGROUNDKeratinocyte 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.
METHODSA 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.
RESULTSMuch 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.
CONCLUSIONK-SFM is an optimal culture medium to maintain and to enrich breast CSCs.
