Isolation, culture and 48 h induction of neural stem cells
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For 48 h induction using serum-free conditions with growth factors, dissociated neurospheres were cultured with recombinant human (rh) BMP4 250 ng/mL , rh Activin A 100 ng/mL and rh bFGF 100 ng/mL individually as well as in combinations as indicated in Supporting Information Fig S1 and
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Growth Factors
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ecombinant human Noggin/Fc chimera and neutralizing anti-human /4 antibody from& , and from.
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Growth Factors
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ecombinant human Noggin/Fc chimera and neutralizing anti-human /4 antibody from& , and from.
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Growth Factors
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ecombinant human Noggin/Fc chimera and neutralizing anti-human /4 antibody from& , and from.
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Effects of Activin A on paraxial mesodermal differentiation of mouse iPS cells.
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The cultures also contained BMP4 (10 ng/ml), IGF-1 (10 ng/ml), LiCl (5 mM), and Shh (10 ng/ml).
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Loss of REX1 within the pluripotent population primes cells for differentiation.
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n = 3 E) Fold enrichment of the percentage of GATA4 positive endoderm cells generated from TRA+VEN− cells relative to those from TRA+VEN+ population after 3 days of treatment with Activin A and BMP4 in low serum media, as observed by immunocytochemistry.
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K02288 selectively inhibits BMP signaling.
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K02288 and LDN-193189 inhibited BMP4 induced Smad1/5/8 phosphorylation in C2C12 cells.
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K02288 selectively inhibits BMP signaling.
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K02288 and LDN-193189 inhibited BMP4 induced Smad1/5/8 phosphorylation in C2C12 cells.
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2.6. Monolayer-Based hiPSC Differentiation towards Definitive Endoderm
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For DE differentiation, hiPSCs were plated onto Matrigel-coated dishes and cultured in medium'>FDTA medium supplemented with 10 μM Rock-inhibitor Y-276342 for 24 hours.
- When the cells reached about 75% confluence, medium was changed to PMI 1640 medium containing 2% FBS with 500 nM IE1 , 3 μM I99021 , 5 μM LY294002 , and 10 ng/mL BMP4 for 24 hours.
- Then, medium was changed to RPMI 1640 medium containing 2% FBS and supplemented with 500 nM IDE1 and 5 μL LY294002 for two days.
- From day 3 on, cells were cultured in RPMI 1640 supplemented with 500 nm IDE1, 5 µM LY294002, and 50 ng/mL FGF2.
- The respective figure contains an experimental outline illustrating detailed culture conditions and treatment regimens [
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Differentiation of ES and iPS Cells
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For definitive endoderm (E) differentiation, mouse ES/iPS cells were cultured on M15 cells with added recombinant human activin-A at 10 ng/ml and/or human bFGF at 5 ng/ml for 3–7 d, as indicated.
- They were subsequently analyzed using flow cytometry to assay for DE or Cer1 expression 5, 0.5 × 105, or 1.0 × 105 cells/well on a matrigel (BD) pre-coated 96-well plate.
- For neuroectoderm differentiation, ES cells were cultured on M15 cells in a differentiation medium supplemented with 10 µM SB431542 (a TGFβ inhibitor)
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