Organ Conditioned Medium: Generating Medium from Harvested Organ Cells for Metastatic Cancer Studies
Published: April 30, 2023
Abstract
Source: Piaseczny, M. M., et al. Generation of Organ-conditioned Media and Applications for Studying Organ-specific Influences on Breast Cancer Metastatic Behavior. J. Vis. Exp.(2016).
This video describes a method of generating an organ conditioned medium derived from the mouse liver. This model system can be used to identify and study organ-derived soluble factors and their effects on the organ tropism and metastatic behavior of cancer cells.
Protocol
1. Generation of Lung- and Brain- Conditioned Media In a sterile tissue culture hood, invert PBS tubes containing lung or brain three times to remove residual blood from organs and aspirate PBS containing blood. Repeat with fresh cold PBS until solution appears clear with no evidence of blood. Place lungs and brains in separate 60 mm2 glass petri dishes. Using two sterile scalpel blades, mince lungs or brains by repeatedly slicing back and forth until tissue fragments are approximat…
Declarações
The authors have nothing to disclose.
Materials
| 1x Phosphate-buffered saline | ThermoFisher Scientific | 10010-023 | Keep sterile |
| 60 mm2 glass petri dishes | Sigma-Aldrich | CLS7016560 | Keep sterile |
| DMEM:F12 | Life Technologies | 21331-020 | Warm in 37 °C water bath before use, keep sterile |
| Penicillin-Streptomycin (10,000 U/ ml) | Life Technologies | 15140-122 | Keep sterile |
| 50 ml conical tubes | Thermo Scientific (Nunc) | 339652 | Keep sterile |
| 0.22 μm syringe filters | Sigma-Aldrich | Z359904 | Keep sterile |
| 6-well tissue culture plates | Thermo Scientific (Nunc) | 140675 | Keep sterile |
| Scalpel blades | Fisher Scientific | S95937A | Keep sterile |
Citar este artigo
Organ Conditioned Medium: Generating Medium from Harvested Organ Cells for Metastatic Cancer Studies. J. Vis. Exp. (Pending Publication), e20218, doi: (2023).