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Found 5 matching solutions for this experiment
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growth media (80% KO-DMEM, 20% Knock-out Serum Replacer (KSR), 1% non-essential amino acids, 2 mM Glutamax, 0.1 mM Ć-Mercaptoethanol, and 1% penicillin and streptomycin all Life Technologies), supplemented with either 10 ng/ml (Shef hES and NAS2 hiPS) or 20 ng/ml (MSU0001) FGF2 (Peprotech). |
Colonies were chopped into uniform 150 Ī¼m pieces using a McIlwain tissue chopper (Mickle Engineering, Gomshall, U.K.), and resuspended in growth media (described above), without FGF2 but supplemented with 10 Ī¼M of the ROCK inhibitor Y27632 (Tocris) |
Colony pieces formed spheres and differentiated into EBs over 4 subsequent days, after which they were transferred into modified chemically defined media (CDM) (Joannides et al., 2007), (BSA was substituted with 400 Ī¼g/ml Albumax-II, Life Technologies). CDM was supplemented with 10 Ī¼M Y27632 (Tocris) and 20 Ī¼M of the Nodal/TGF-Ī² signaling inhibitor SB431542 (Tocris) |
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growth media (80% KO-DMEM, 20% Knock-out Serum Replacer (KSR), 1% non-essential amino acids, 2 mM Glutamax, 0.1 mM Ć-Mercaptoethanol, and 1% penicillin and streptomycin all Life Technologies), supplemented with either 10 ng/ml (Shef hES and NAS2 hiPS) or 20 ng/ml (MSU0001) FGF2 (Peprotech). |
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Colonies were chopped into uniform 150 Ī¼m pieces using a McIlwain tissue chopper (Mickle Engineering, Gomshall, U.K.), and resuspended in growth media (described above), without FGF2 but supplemented with 10 Ī¼M of the ROCK inhibitor Y27632 (Tocris) |
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Colony pieces formed spheres and differentiated into EBs over 4 subsequent days, after which they were transferred into modified chemically defined media (CDM) (Joannides et al., 2007), (BSA was substituted with 400 Ī¼g/ml Albumax-II, Life Technologies). CDM was supplemented with 10 Ī¼M Y27632 (Tocris) and 20 Ī¼M of the Nodal/TGF-Ī² signaling inhibitor SB431542 (Tocris) |
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Growth Medium: DMEM/F12 (Merck Millipore, Germany) supplemented with 20% KnockOutā¢ Serum Replacement (KnockOutā¢ SR, Thermo Fisher Scientific Inc., MA, USA), 1% penicillin-streptomycin (Sigma-Aldrich; Merck KGaA), 1 mM non-essential amino acid (NEAA; Sigma-Aldrich; Merck KGaA), 0.2 mM 2-mercaptoethanol (Sigma-Aldrich; Merck KGaA), and 10 ng/ml basic fibroblast growth factor (bFGF) (Sigma-Aldrich; Merck KGaA) |
Growth medium with the addition of 10 ĀµM ROCK-inhibitor Y-27632 (Sigma-Aldrich; Merck KGaA). Next, cells were left for spontaneous aggregation for 15 days. The medium (hESC medium without bFGF) was changed every second day. |
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Growth Medium: DMEM/F12 (Merck Millipore, Germany) supplemented with 20% KnockOutā¢ Serum Replacement (KnockOutā¢ SR, Thermo Fisher Scientific Inc., MA, USA), 1% penicillin-streptomycin (Sigma-Aldrich; Merck KGaA), 1 mM non-essential amino acid (NEAA; Sigma-Aldrich; Merck KGaA), 0.2 mM 2-mercaptoethanol (Sigma-Aldrich; Merck KGaA), and 10 ng/ml basic fibroblast growth factor (bFGF) (Sigma-Aldrich; Merck KGaA) |
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Growth medium with the addition of 10 ĀµM ROCK-inhibitor Y-27632 (Sigma-Aldrich; Merck KGaA). Next, cells were left for spontaneous aggregation for 15 days. The medium (hESC medium without bFGF) was changed every second day. |
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human pluripotent stem cells were detached from the feeder layer by collagenase IV, dispersed into small clumps |
Iscoveās modified Dulbeccoās differentiation medium supplemented with 20% fetal calf serum, 1Ā mMĀ l-glutamine, 0.1Ā mM beta-mercaptoethanol and 1% nonessential amino acid stock in ultra-low attachment plates (Corning) for 7Ā days. |
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human pluripotent stem cells were detached from the feeder layer by collagenase IV, dispersed into small clumps |
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Iscoveās modified Dulbeccoās differentiation medium supplemented with 20% fetal calf serum, 1Ā mMĀ l-glutamine, 0.1Ā mM beta-mercaptoethanol and 1% nonessential amino acid stock in ultra-low attachment plates (Corning) for 7Ā days. |
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The cells were centrifuged at 300g for 3 min and then resuspended in Apelā¢ media (Stem Cell Technologies) containing 10 Ī¼M Rock inhibitor at 30,000 cells per ml. 100 Ī¼l was then added to each well of a U-bottom 96 well plate and centrifuged at 300g for 3 min. |
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The cells were centrifuged at 300g for 3 min and then resuspended in Apelā¢ media (Stem Cell Technologies) containing 10 Ī¼M Rock inhibitor at 30,000 cells per ml. 100 Ī¼l was then added to each well of a U-bottom 96 well plate and centrifuged at 300g for 3 min. |
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Cell aggregates were suspended in EB formation medium (50% DMEM/F12 (Life Technologies, 11330-032) 50% mTeSRā¢1 or StemProĀ® hESC SFM containing 10 ĀµM Rock Inhibitor Y27632 (Millipore, SCM075)) and allowed to settle by gravity in a conical tube. |
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Cell aggregates were suspended in EB formation medium (50% DMEM/F12 (Life Technologies, 11330-032) 50% mTeSRā¢1 or StemProĀ® hESC SFM containing 10 ĀµM Rock Inhibitor Y27632 (Millipore, SCM075)) and allowed to settle by gravity in a conical tube. |
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