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Matrigel-based multi-spheroid 3D growth assays

HA Hussein Al-Akhrass
JC James R. W. Conway
AP Annemarie Svane Aavild Poulsen
IP Ilkka Paatero
JK Jasmin Kaivola
AP Artur Padzik
OA Olav M. Andersen
JI Johanna Ivaska
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The bottom wells of a µ-Plate 96 well plate (ibidi, 89646) were filled with 10 µL of 50% matrigel (Corning, 356231) then the plate was centrifuged at 200 × g for 20 min. The coated plate was incubated at 37 °C for 30 min. Wells were then filled with 20 µL of cell suspension (1000 cells, 1:1 ratio of BT-474 cells to TIFF) in 25% matrigel and incubated overnight at 37 °C. Wells were then filled with 65 µL of cell culture medium that was replaced every 2 days. Wells were imaged using the IncuCyte® S3 instrument (sartorius) and spheroid growth, reflected by mCherry fluorescence, was analyzed using the IncuCyte software (sartorious).

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