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Found 2 matching solutions for this experiment
Upstream tips |
Protocol tips |
Downstream tips |
In matrigel, Y-27632 (10 μM) was included for the first 2 days to avoid anoikis |
Advanced DMEM/F12 supplemented with Penicillin/Streptomycin, 10 mM Hepes, Glutamax, 1× N2, 1× B27 (all from Invitrogen), and 1 μM N-acetylcysteine (Sigma) containing growth factors: 50 ng/ml EGF, 100 ng/ml noggin, 1 μg/ml R-spondin) were overlaid |
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Upstream tips |
In matrigel, Y-27632 (10 μM) was included for the first 2 days to avoid anoikis |
Protocol tips |
Advanced DMEM/F12 supplemented with Penicillin/Streptomycin, 10 mM Hepes, Glutamax, 1× N2, 1× B27 (all from Invitrogen), and 1 μM N-acetylcysteine (Sigma) containing growth factors: 50 ng/ml EGF, 100 ng/ml noggin, 1 μg/ml R-spondin) were overlaid |
Upstream tips |
Protocol tips |
Downstream tips |
Pelleted crypt structures were resuspended in a 1:1 mixture of Intesticult medium (Stemcell technologies, Köln, Germany) and growth factor reduced matrigel (Corning, NY, USA) |
Intesticult medium supplemented with 100 U/ml Penicillin, 100 μg/ml Streptomycin, 2.5 μg/ml amphotericin B, 2.05 mg/ml sodium deoxycholate (all Thermo Fisher scientific) and 20 μg/ml Gentamycin (Lonza, Basel Switzerland) was added (300 μl/well) |
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Upstream tips |
Pelleted crypt structures were resuspended in a 1:1 mixture of Intesticult medium (Stemcell technologies, Köln, Germany) and growth factor reduced matrigel (Corning, NY, USA) |
Protocol tips |
Intesticult medium supplemented with 100 U/ml Penicillin, 100 μg/ml Streptomycin, 2.5 μg/ml amphotericin B, 2.05 mg/ml sodium deoxycholate (all Thermo Fisher scientific) and 20 μg/ml Gentamycin (Lonza, Basel Switzerland) was added (300 μl/well) |
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