NMP differentiation was based on the published protocols20 (link),56 (link) with minor modifications. In preparation for differentiation, ESCs were feeder-depleted and plated in gelatin-coated (0.1% Sigma, G1890-100G) 6-well plates (Falcon, 353046) at a density of 8×103 cells/cm2 in ES medium. On D0 of differentiation media was changed to N2B27 media (Composition: 49.5% Advanced Dulbecco’s Modified Medium F-12 (Gibco, 12634028); 49% Neurobasal medium (Gibco, 21103049); 0.5% N2-supplement (Gibco, 17502001); 1% B27-supplement (Gibco, 17504044)) supplied with 1x Glutamax (Gibco, 17504044), 40 µg/ml BSA Fraction V (Gibco, 15260037) and 100 mM 2-Mercaptoethanol (Gibco, 21985-023)) supplemented with 10 ng/ml hFGF-2 (Miltenyi Biotec, 130-104-925). The media was further supplemented with 5 µM CHIR99021 (StemMACS, 130-103-926) on D2 and with or without 50 ng/ml hGdf11 (130-105-776, Miltenyi Biotec) on D3. Throughout differentiation, the medium was refreshed every 24 h. NMP identity at D3 was routinely confirmed by co-expression analysis of Sox2 and T/Bra using immunofluorescence.
Hgdf11
Manufactured by Miltenyi Biotec
The HGdf11 is a laboratory equipment product offered by Miltenyi Biotec. It is a device used in various applications within the field of biotechnology. The core function of the HGdf11 is to perform specific tasks required in research and analysis procedures, but a detailed description of its intended use cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Bsa fraction 5, by Thermo Fisher Scientific (2 mentions)
B27 supplement, by Thermo Fisher Scientific (2 mentions)
Hfgf 2, by Miltenyi Biotec (2 mentions)
N2 supplement, by Thermo Fisher Scientific (2 mentions)
2 mercaptoethanol, by Thermo Fisher Scientific (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
Neurobasal medium, by Thermo Fisher Scientific (2 mentions)
2 protocols using hgdf11
1
NMP Differentiation from Embryonic Stem Cells
2
NMP Differentiation from Embryonic Stem Cells
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NMP differentiation was based on the published protocols (Gouti et al., 2014 (Gouti et al., , 2017) ) with minor modifications. In preparation for differentiation, ESCs were feeder-depleted and plated in gelatin-coated (0.1% Sigma, G1890-100G) 6-well plates (Falcon, 353046) at a density of 8x10 3 cells/cm 2 in ES medium. On D0 of differentiation media was changed to N2B27 media -12 (Gibco, 12634028); 49%
Neurobasal medium (Gibco, 21103049); 0.5% N2-supplement (Gibco, 17502001); 1% B27supplement (Gibco, 17504044)) supplied with 1x Glutamax (Gibco, 17504044), 40 µg/ml BSA Fraction V (Gibco, 15260037) and 100 mM 2-Mercaptoethanol (Gibco, 21985-023))
supplemented with 10 ng/ml hFGF-2 (Miltenyi Biotec, . The media was further supplemented with 5µM CHIR99021 (StemMACS, 130-103-926) on D2 and with or without 50 ng/ml hGdf11 (130-105-776, Miltenyi Biotec) on D3. Throughout differentiation, the medium was refreshed every 24 hours. NMP identity at D3 was routinely confirmed by coexpression analysis of Sox2 and T/Bra using immunofluorescence.
Neurobasal medium (Gibco, 21103049); 0.5% N2-supplement (Gibco, 17502001); 1% B27supplement (Gibco, 17504044)) supplied with 1x Glutamax (Gibco, 17504044), 40 µg/ml BSA Fraction V (Gibco, 15260037) and 100 mM 2-Mercaptoethanol (Gibco, 21985-023))
supplemented with 10 ng/ml hFGF-2 (Miltenyi Biotec, . The media was further supplemented with 5µM CHIR99021 (StemMACS, 130-103-926) on D2 and with or without 50 ng/ml hGdf11 (130-105-776, Miltenyi Biotec) on D3. Throughout differentiation, the medium was refreshed every 24 hours. NMP identity at D3 was routinely confirmed by coexpression analysis of Sox2 and T/Bra using immunofluorescence.
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