Mouse breast cancer cell (4T1) and colon cancer cell (CT26) were purchased from ATCC (Manassas, VA, USA) and cultured in RPMI-1640 complete medium containing 10% fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, USA). Both cell lines were cultivated with 5% at 37 °C CO2 in a humidified atmosphere and treated with G-CSF (Shenandoah Biotechnology, Warwick, PA) for either 10 days (short term) or 7 weeks (long term) at 25 ng/ml. Our previous study 20 (link) showed that the cytokine expressions after 6 weeks treatment of G-CSF were significantly different with short term groups. So we selected 7 weeks treatment in this study. Cells treated with PBS were used as the control group. Supernatants of long-term culture were saved for cytokine analysis and cell pellets were saved from RNA extraction.
Protocol detail
Long-term G-CSF Exposure in Breast and Colon Cancer
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Atmosphere
Breast cancer
Cell
Cell lines
Colon cancer
Cultured medium
Cytokine
Fetal bovine serum
G csf
Mouse
Pellets
Corresponding Organization : University of Utah
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Variable analysis
independent variables
- G-CSF treatment duration (short term - 10 days, long term - 7 weeks)
- Cytokine expressions
- Cell pellets for RNA extraction
- Cell lines (mouse breast cancer cell 4T1 and mouse colon cancer cell CT26)
- Cell culture medium (RPMI-1640 complete medium containing 10% fetal bovine serum)
- Cell culture conditions (5% CO2, 37°C, humidified atmosphere)
- Positive control: Cells treated with G-CSF (25 ng/ml)
- Negative control: Cells treated with PBS
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