3D Co-culture with Direct Interaction: Co-culturing Cancer Cells with Monocytes to Study Their Interaction
Published: April 30, 2023
Abstract
Source: Espinoza-Sánchez, et al. Analyzing the Communication Between Monocytes and Primary Breast Cancer Cells in an Extracellular Matrix Extract (ECME)-based Three-dimensional System. J. Vis. Exp. (2018).
This video describes the 3D co-culture of breast cancer cells with monocytes to study their interaction in an extracellular matrix extract (ECME) based environment. This method helps in studying specific features such as collagen degradation, immune cell recruitment, and cell invasion.
Protocol
1. 3D co-cultures with direct interaction Label BrC cells and monocytes with different fluorescent dyes before cell culture to allow independent sorting of each cell lineage after culture for analysis of mRNA and protein expression. Use commercially available derivatives of coumarin and rhodamine that freely pass through the cell membrane of living cells. A general protocol for labeling is the following: Prepare a monolayer of BrC cells of interest (2 × 106 ce…
Declarações
The authors have nothing to disclose.
Materials
| U937 | American Type Culture Collection ATCC | CRL-1593.2 | Monocytic cell line/histiocytic lymphoma |
| THP-1 | American Type Culture Collection ATCC | TIB-202 | Monocytic cell line/acute monocytic leukemia |
| MCF-10A | American Type Culture Collection ATCC | CRL-10317 | Non-transformed breast cell line |
| MCF-7 | American Type Culture Collection ATCC | HTB-22 | Breast Cancer Cell line |
| MDA-MB-231 | American Type Culture Collection ATCC | HTB-26 | Breast Cancer Cell line |
| RPMI 1640 medium | GIBCO BRL Life Technologies | 11875-093 | |
| DMEM/F12 | GIBCO BRL Life Technologies | 11039-021 | |
| Antibiotic/Antimycotic | GIBCO BRL Life Technologies | 15240-062 | 100 U/mL penicillin, 100 µg/mL streptomycin, and 0.25 µg/mL Fungizone |
| Fetal Bovine Serum | GIBCO BRL Life Technologies | 16000-044 | |
| Horse serum | GIBCO BRL Life Technologies | 16050114 | |
| 0.05% Trypsin-EDTA 1X | GIBCO BRL Life Technologies | 25300-062 | |
| PBS 1X (Phosphate Buffered Saline | GIBCO BRL Life Technologies | 20012-027 | |
| Epidermal Growth Factor (EGF) | PeproTech | AF-100-15 (1.00mg) | |
| Insulin | SIGMA-ALDRICH | I1882-100MG | |
| Hydrocortisone | SIGMA-ALDRICH | H088-5G | |
| Cholera toxin Vibrio cholerae | SIGMA-ALDRICH | C8052-1MG | |
| Matrigel | Corning Inc | 356237 | Engelbreth-Holm-Swarm (EHS) mouse sarcoma, extracellular matrix extract, Store at -20°C until use at 4°C |
| Lab-Tek chamber slide with cover (8 well) | Nalge Nunc International | 177402 |
Citar este artigo
3D Co-culture with Direct Interaction: Co-culturing Cancer Cells with Monocytes to Study Their Interaction. J. Vis. Exp. (Pending Publication), e20216, doi: (2023).