Organoid Culture Conditions for hM1A and hT2

Organoids (hM1A and hT2) were kindly provided by the Tuveson laboratory (Cold Spring Harbor Laboratory). Organoids were cultured at 37°C in 5% CO₂. Cells were seeded in growth factor-reduced Matrigel (Corning; Cat# 356231) domes and fed with human complete feeding medium: advanced DMEM/F12-based WRN-conditioned medium (L-WRN (ATCC CRL-3276)), 1x B27 supplement, 10 mM HEPES, 0.01 μM GlutaMAX, 10 mM nicotinamide, 1.25 mM N-acetylcysteine, 50 ng/mL hEGF, 100 ng/mL hFGF10, 0.01 μM hGastrin I, 500 nM A83–01, 1 μM PGE2 and 10.5 μM Y27632 (44 (link)).

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Hobbs G.A., Baker N.M., Miermont A.M., Thurman R.D., Pierobon M., Tran T.H., Anderson A.O., Waters A.M., Diehl J.N., Papke B., Hodge R.G., Klomp J.E., Goodwin C.M., DeLiberty J.M., Wang J., Ng R.W., Gautam P., Bryant K.L., Esposito D., Campbell S.L., Petricoin EF I.I.I., Simanshu D.K., Aguirre A.J., Wolpin B.M., Wennerberg K., Rudloff U., Cox A.D, & Der C.J. (2019). Atypical KRASG12R Mutant Is Impaired in PI3K Signaling and Macropinocytosis in Pancreatic Cancer. Cancer discovery, 10(1), 104-123.

Publication 2019

Matrigel L-WRN

A83 01 Acetylcysteine Cells Cold Conditioned medium Growth factor Hepes Human Matrigel Nicotinamide Organoids Pge2 Supplement Y27632

Corresponding Organization : University of North Carolina at Chapel Hill

Other organizations : George Mason University, Frederick National Laboratory for Cancer Research, Dana-Farber Cancer Institute, Institute for Molecular Medicine Finland, University of Helsinki, Broad Institute, University of Copenhagen

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independent variables

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dependent variables

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control variables

Cells were cultured at 37°C in 5% CO2
Cells were seeded in growth factor-reduced Matrigel (Corning; Cat# 356231) domes
Cells were fed with human complete feeding medium: advanced DMEM/F12-based WRN-conditioned medium (L-WRN (ATCC CRL-3276)), 1x B27 supplement, 10 mM HEPES, 0.01 μM GlutaMAX, 10 mM nicotinamide, 1.25 mM N-acetylcysteine, 50 ng/mL hEGF, 100 ng/mL hFGF10, 0.01 μM hGastrin I, 500 nM A83–01, 1 μM PGE2 and 10.5 μM Y27632

controls

No positive or negative controls were explicitly mentioned in the provided information.

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