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.
Hfgf 2
Manufactured by Miltenyi Biotec
The HFGF-2 is a recombinant human fibroblast growth factor-2 (FGF-2) product developed by Miltenyi Biotec. FGF-2 is a protein that plays a role in cell proliferation, differentiation, and survival.
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)
Hgdf11, 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)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Testosterone, by Merck Group (1 mentions)
3 protocols using hfgf 2
1
NMP Differentiation from Embryonic Stem Cells
2
Primary Prostate Fibroblast Cultures
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fresh patient tissues were collected from radical prostatectomy specimens with written informed consent from patients. These studies were approved by Human Research Ethics Committees at Monash University (2004/145) and Cabrini Hospital (03-14-04-08) in Melbourne, Australia. Primary cultures of CAFs were established from tumor tissue and matched NPFs from non-malignant tissue as previously described [36 (link)]. A board-certified pathologist confirmed that the tumor tissue contained approximately 80% cancer and that the non-malignant tissue was benign. Cells were maintained in RPMI 1640 media (no phenol red; Gibco, ThermoFisher) supplemented 5% fetal bovine serum (FBS; Gibco, ThermoFisher), 1 nM testosterone (Sigma-Aldrich), 10 ng/mL human fibroblast growth factor 2 (hFGF-2; Miltenyi Biotec), 100 U/mL penicillin, and 100 μg/mL streptomycin (Gibco, ThermoFisher). Cells were maintained at 37 °C in 5% CO2, with media changes every 2–3 days. Matched fibroblasts from two patients were used for this study between passages 5 and 8.
3
NMP Differentiation from Embryonic Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!