In vitro viability of 4T1-Luc2-tdTomato cancer cells after co-culturing with cells derived from D-lung or D-scaffold

JW Jing Wang
RO Ramon Ocadiz-Ruiz
MH Matthew S. Hall
GB Grace G. Bushnell
SO Sophia M. Orbach
JD Joseph T. Decker
RR Ravi M. Raghani
YZ Yining Zhang
AM Aaron H. Morris
JJ Jacqueline S. Jeruss
LS Lonnie D. Shea
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4T1-Luc2-tdTomato cancer cells were seeded into 96-well plate at a density of 800 cells per 200 µl per well in complete media supplemented with 0.6 mM D-luciferin, and then incubated at 37 °C overnight to allow attachment. The next day, scaffolds and lungs were retrieved from 4T1-bearing BALB/c mice at two weeks after tumor inoculation and prepared to single-cell suspensions. Scaffold or lung cells were added to 4T1-Luc2-tdTomato cell cultures in D-luciferin-included complete media at the desired cancer cell to niche cell ratios, and then co-cultured for two days at 37 °C. The number of viable cancer cells was quantified by measuring the bioluminescence of 4T1-Luc2-tdTomato cancer cells using a plate reader (Synergy H1 hybrid multi-mode reader, BioTek).

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