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Found 3 matching solutions for this experiment
Upstream tips |
Protocol tips |
Downstream tips |
20 000 cells per 200 μl in a V-shaped well of a 96-well plate (Sumitomo Bakelite, Tokyo, Japan) with an ultra-low-cell adhesion surface |
KnockOut DMEM (Life Technologies, Carlsbad, CA, USA) containing 15% KnockOut Serum Replacement (HyClone, Logan, UT, USA), 1% GlutaMAX (Life Technologies), 1% MEM Non-Essential Amino Acids (Life Technologies), 0.09% 2-mercaptoethanol (Life Technologies), and 0.1% Leukemia Inhibitory Factor (Wako, Osaka, Japan) |
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Upstream tips |
20 000 cells per 200 μl in a V-shaped well of a 96-well plate (Sumitomo Bakelite, Tokyo, Japan) with an ultra-low-cell adhesion surface |
Protocol tips |
KnockOut DMEM (Life Technologies, Carlsbad, CA, USA) containing 15% KnockOut Serum Replacement (HyClone, Logan, UT, USA), 1% GlutaMAX (Life Technologies), 1% MEM Non-Essential Amino Acids (Life Technologies), 0.09% 2-mercaptoethanol (Life Technologies), and 0.1% Leukemia Inhibitory Factor (Wako, Osaka, Japan) |
Upstream tips |
Protocol tips |
Downstream tips |
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differentiation medium (G-MEM (Gibco #11710-035), supplemented with 10% KnockOut™ Serum Replacement (Gibco #10828-028, Lot 1667387), 1% non-essential amino acids (GE Healthcare), 0.1 mM β-mercaptoethanol (Sigma Aldrich), 200 U/ml penicillin (Sigma Aldrich), 200 μg/ml streptomycin (Sigma Aldrich)) per 1/96 well (Nunclon Sphera® Low Adhesion Plate 96-U, Thermo Scientific #174925). |
On day six, aggregates were transferred to 90 mm plates with low adhesion surface (Nunclon Sphera® 90 mm, Thermo Scientific #174945) and subsequently cultured in N2 medium (49% DMEM low glucose (1 g/l, GE Healthcare #E15-005), 49% HAM's F12 (GE Healthcare # E15-016), 1 × N2 supplement (100 ×, Gibco #17502-048 Lot 1737260) and 200 U/ml penicillin (Sigma Aldrich), 200 μg/ml streptomycin (Sigma Aldrich)). |
Protocol tips |
differentiation medium (G-MEM (Gibco #11710-035), supplemented with 10% KnockOut™ Serum Replacement (Gibco #10828-028, Lot 1667387), 1% non-essential amino acids (GE Healthcare), 0.1 mM β-mercaptoethanol (Sigma Aldrich), 200 U/ml penicillin (Sigma Aldrich), 200 μg/ml streptomycin (Sigma Aldrich)) per 1/96 well (Nunclon Sphera® Low Adhesion Plate 96-U, Thermo Scientific #174925). |
Downstream tips |
On day six, aggregates were transferred to 90 mm plates with low adhesion surface (Nunclon Sphera® 90 mm, Thermo Scientific #174945) and subsequently cultured in N2 medium (49% DMEM low glucose (1 g/l, GE Healthcare #E15-005), 49% HAM's F12 (GE Healthcare # E15-016), 1 × N2 supplement (100 ×, Gibco #17502-048 Lot 1737260) and 200 U/ml penicillin (Sigma Aldrich), 200 μg/ml streptomycin (Sigma Aldrich)). |
Upstream tips |
Protocol tips |
Downstream tips |
cells were transferred to low adhesion tissue culture plates coated with 2% poly-HEMA in ethanol. |
EBM [DMEM-F12, 15% Knock Out Serum Replacement (KOSR), 15% HI FBS, 1 mM L-glutamine, 0.1 mM 2-mercaptoethanol, and 1% Non-essential amino acids (NEAA, Invitrogen, Grand Island, NY), 1% antibiotic-antimycotic solution (Invitrogen), and 40 ng/ml human Beta fibroblast growth factor (β-FGF)] in suspension for a minimum of 14 days, during which they formed EBs. |
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Upstream tips |
cells were transferred to low adhesion tissue culture plates coated with 2% poly-HEMA in ethanol. |
Protocol tips |
EBM [DMEM-F12, 15% Knock Out Serum Replacement (KOSR), 15% HI FBS, 1 mM L-glutamine, 0.1 mM 2-mercaptoethanol, and 1% Non-essential amino acids (NEAA, Invitrogen, Grand Island, NY), 1% antibiotic-antimycotic solution (Invitrogen), and 40 ng/ml human Beta fibroblast growth factor (β-FGF)] in suspension for a minimum of 14 days, during which they formed EBs. |
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