No discussions found
Start your discussion
Share your thoughts or question with experts in your field
Start a discussion
Found 3 matching solutions for this experiment
Upstream tips |
Protocol tips |
Downstream tips |
|
DMEM/F12-GLUTAMAX-type medium containing 4% B27 supplement without vitamin A (Invitrogen), 1% N2 supplement (Invitrogen), and 55 ďM 2-mercaptoethanol (2-ME). To induce forebrain fate, the medium was supplemented with 5ÂľM Y-27632 and 200ng/ml recombinant mouse Noggin (R&D Systems, 1967-NG-025) |
After 2 days, EBs were collected and plated onto 10-cm bacterial Petri dishes in 10 ml neuronal medium as above with the addition of B27 supplemented with vitamin A. |
Protocol tips |
DMEM/F12-GLUTAMAX-type medium containing 4% B27 supplement without vitamin A (Invitrogen), 1% N2 supplement (Invitrogen), and 55 ďM 2-mercaptoethanol (2-ME). To induce forebrain fate, the medium was supplemented with 5ÂľM Y-27632 and 200ng/ml recombinant mouse Noggin (R&D Systems, 1967-NG-025) |
Downstream tips |
After 2 days, EBs were collected and plated onto 10-cm bacterial Petri dishes in 10 ml neuronal medium as above with the addition of B27 supplemented with vitamin A. |
Upstream tips |
Protocol tips |
Downstream tips |
|
increasing amounts of N2 media (25, 50, 75, 100%) were added to the mTeSR medium every other day whilst maintaining 2.5uM dorsomorphin. N2 medium contains 1X N2 supplement (Invitrogen), 50% DMEM/F12, 50% Neurobasal, 0.075% Bovine serum albumin, and 2mM GlutaMAX (Life Technologies) |
Cells were subsequently passaged with trypsin and cultured on matrigel-coated dishes in N2 media with the addition of 20ng/ml bFGF. Rho-associated protein kinase (ROCK) inhibitor Y27632 (10uM, Stemgent) was added to the medium for the first 24 hours. NPs were expanded and maintained in N2 medium with 20ng/ml bFGF. All 2D culture experiments were performed on WIBR3-derived NPs unless otherwise specified. |
Protocol tips |
increasing amounts of N2 media (25, 50, 75, 100%) were added to the mTeSR medium every other day whilst maintaining 2.5uM dorsomorphin. N2 medium contains 1X N2 supplement (Invitrogen), 50% DMEM/F12, 50% Neurobasal, 0.075% Bovine serum albumin, and 2mM GlutaMAX (Life Technologies) |
Downstream tips |
Cells were subsequently passaged with trypsin and cultured on matrigel-coated dishes in N2 media with the addition of 20ng/ml bFGF. Rho-associated protein kinase (ROCK) inhibitor Y27632 (10uM, Stemgent) was added to the medium for the first 24 hours. NPs were expanded and maintained in N2 medium with 20ng/ml bFGF. All 2D culture experiments were performed on WIBR3-derived NPs unless otherwise specified. |
Upstream tips |
Protocol tips |
Downstream tips |
NS cells were grown on poly-L-ornithin and laminin coated plates. Poly-L-ornithine/laminin plates were generated as such: A 20 ug/ml solution of poly-L-ornithine (Sigma) in water was added to plates and plates were incubated at 37°C for 1 hour. The poly-L-ornithine solution was then removed, plates were washed three times with PBS, and then 5 ul/ml solution of laminin (Sigma) in PBS was added to plates and plates were incubated at 37°C for at least 3 hours. |
1:1 mix of DMEM/F12:Neurobasal (Invitrogen), 0.5X N2 (Invitrogen), 0.5X B27 (Invitrogen), 1X Glutamax, and 0.1 mM beta-mercaptoethanol, which was supplemented with 20 ng/ul of both EGF and FGF-2 (R&D) |
|
Upstream tips |
NS cells were grown on poly-L-ornithin and laminin coated plates. Poly-L-ornithine/laminin plates were generated as such: A 20 ug/ml solution of poly-L-ornithine (Sigma) in water was added to plates and plates were incubated at 37°C for 1 hour. The poly-L-ornithine solution was then removed, plates were washed three times with PBS, and then 5 ul/ml solution of laminin (Sigma) in PBS was added to plates and plates were incubated at 37°C for at least 3 hours. |
Protocol tips |
1:1 mix of DMEM/F12:Neurobasal (Invitrogen), 0.5X N2 (Invitrogen), 0.5X B27 (Invitrogen), 1X Glutamax, and 0.1 mM beta-mercaptoethanol, which was supplemented with 20 ng/ul of both EGF and FGF-2 (R&D) |
Can't find the product you've used to perform this experiment? It would be great if you can help us by
Adding a product!