Mobile website
Basic Research
Enrichment and identification of cancer stem cell-like cells in mouse breast cancer cell line 4T1
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective:To culture mouse breast cancer 4T1 cells in serum-free medium(SFM), and to screen for and identify the cancer stem-like cells in 4T1 cells. Methods: Breast cancer stem-like cells were enriched from 4T1 cells cultured in SFM containing EGF, bFEGF, and B27, etc. The suspension spheres were seeded in serum-supplement medium (SSM) and cell differentiation was observed. The proportion of cancer stem-like cells in 4T1 cells was determined through cell surface markers CD44+CD24-/low and Hoechst 33342 staining. Tumorigenic abilities of 4T1 cells in different culture conditions were detected by mouse tumorgenesis experiment. Results: 4T1 cells could survive, proliferate and form breast cancer suspension spheres in SFM for prolonged time period. 4T1 spheres seeded into SSM could differentiate and adhere to the culture plates. There were 6.4%-68.9% CD44+CD24-/low cells and 7.3%-61.2% side population (SP) cells in 4T1 spheres, which were significantly higher than those in 4T1 cells cultured in SSM (P<0.05). The ratios of CD44_CD24-/low and SP cells were gradually increased with the passage of 4T1 spheres in SFM. 4T1 spheres with enriched cancer stem cell were more tumorigenic than 4T1 cells cultured in the SSM. Conclusion: 4T1 cells can grow and form spheres in serum-free suspension medium containing growth factor, and they contain breast cancer stem-like cells, which can be enriched when cultured in SFM.
Keywords:
Project Supported:
Project supported by the Natural Science Foundation of Hebei Province (No. C2009001195)
Clc Number:
