Estrogen-suppressive organoids were established from fresh surgical specimens of GS3 using 3D culture conditions (Fig. S2). After tumor tissue was minced and digested, 10 4 of organoids were embedded in VitroGel 3D-RGD (TWG002, TheWell Bioscience, North Brunswick, NJ) on a 96 well plate and cultured in E2-free M87 medium at 37˚C [23] . The organoids viability was evaluated using the CellTider-Glo 3D Cell Viability Assay (G9682, Promega, Madison, WI) which measured ATP. We made at least ve technical replicates and repeat experiments twice.
Celltider glo 3d cell viability assay
Manufactured by Promega
The CellTiter-Glo 3D Cell Viability Assay is a luminescent cell viability assay that quantifies ATP as an indicator of metabolically active cells. The assay is designed for use with 3D cell culture models.
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2 protocols using celltider glo 3d cell viability assay
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Estrogen-Suppressed Tumor Organoid Cultivation
2
Establishing E2-Suppressing Tumor Organoids
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Organoids with the E2-suppressing phenotype were established from PDX-derived surgical specimens of GS3 tumors using 3D culture conditions (Fig. S3). After tumor tissue was minced and digested, 10 4 organoids were embedded in VitroGel 3D-RGD (TWG002, TheWell Bioscience, North Brunswick, NJ) on a 96 well plate and cultured in E2-free M87 medium at 37˚C [13] (link). Organoids viability was evaluated using the CellTider-Glo 3D Cell Viability Assay (G9682, Promega, Madison, WI) which measured ATP. We made at least ve technical replicates and repeated each experiment twice.
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