To obtain neuronal spheroids, hBM-MSCs were cultured in a low-attachment dish (Corning) with serum-free sphere-forming medium comprising advanced DMEM/F-12 (1:1) (Gibco) supplemented with 0.5 mM l-glutamine, N2 supplement (Gibco), 20 ng/mL recombinant human EGF (Wako), 20 ng/mL human FGF-2 (Wako), and 2% B27 (Gibco). Following the formation of spheroids, neuronal spheroids obtained using low-attachment dishes or MSC spheroids obtained under shaking-culture conditions were seeded on fibronectin (Wako)-coated 6-well chambers (Matsunami Glass Ind. Inc., Kishiwada, Osaka, Japan) using induction medium comprising advanced DMEM/F12 (1:1) (Gibco) supplemented with 10% FBS (Hyclone, GE Healthcare), N2 supplement (Gibco), 1% P/S (Wako), and 10 mM HEPES (Dojindo Molecular Technologies, Inc.) (Morikawa et al., 2009b (link); Choi et al., 2017 (link)). The medium was changed every 3–4 days until day 10.
Human fgf 2
Manufactured by Fujifilm
Sourced in Japan
Human FGF-2 is a recombinant protein that represents the fibroblast growth factor 2 (FGF-2) found in humans. FGF-2 is a member of the fibroblast growth factor family and plays a role in various cellular processes, including cell growth, differentiation, and angiogenesis. The product is intended for use in research and laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
Advanced dmem f12, by Thermo Fisher Scientific (1 mentions)
N2 supplement, by Thermo Fisher Scientific (1 mentions)
Hepes, by Dojindo Laboratories (1 mentions)
Recombinant human egf, by Fujifilm (1 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
Nucleoside, by Merck Group (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Es qualified fbs, by Thermo Fisher Scientific (1 mentions)
Monothioglycerol, by Fujifilm (1 mentions)
2 protocols using human fgf 2
1
Neuronal Spheroid Formation from hBM-MSCs
2
Isolation and Culture of Chick Primordial Germ Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Circulating PGCs were collected from male WL embryonic blood at Hamburger and Hamilton’s developmental stage
14 (HH14) [25 (link)]. Embryos were sexed by PCR using W chromosome-specific
primers [26 (link)], and whole blood cells containing PGCs were grown on a BRL
feeder layer. The PGC culture medium was prepared as described in previous studies with some modifications
[1 (link), 5 ]. The PGC culture medium
contained KnockOut DMEM (KO-DMEM; Life Technologies), 50% BRL-conditioned KO-DMEM, 7.5% ES qualified FBS (Life
Technologies), 2.5% chicken serum (Biowest, Nuaillé, France), 2 mM GlutaMAX (Life Technologies), 1 mM sodium
pyruvate (Life Technologies), 1 × nonessential amino acids (Life Technologies), 1 × nucleosides (Millipore,
Billerica, MA, USA), 0.5 mM monothioglycerol (Wako Pure Chemical Industries, Osaka, Japan) and 5 ng/ml human
FGF2 (Wako Pure Chemical Industries). Half the volume of the culture medium was replaced every 2 days during
the culture period for PGCs, and the cultured PGCs were passaged until 90% confluence. Several male PGCs were
proliferated to large numbers, and could be maintained for more than 100 days. The established PGC lines were
used for subsequent assays.
14 (HH14) [25 (link)]. Embryos were sexed by PCR using W chromosome-specific
primers [26 (link)], and whole blood cells containing PGCs were grown on a BRL
feeder layer. The PGC culture medium was prepared as described in previous studies with some modifications
[1 (link), 5 ]. The PGC culture medium
contained KnockOut DMEM (KO-DMEM; Life Technologies), 50% BRL-conditioned KO-DMEM, 7.5% ES qualified FBS (Life
Technologies), 2.5% chicken serum (Biowest, Nuaillé, France), 2 mM GlutaMAX (Life Technologies), 1 mM sodium
pyruvate (Life Technologies), 1 × nonessential amino acids (Life Technologies), 1 × nucleosides (Millipore,
Billerica, MA, USA), 0.5 mM monothioglycerol (Wako Pure Chemical Industries, Osaka, Japan) and 5 ng/ml human
FGF2 (Wako Pure Chemical Industries). Half the volume of the culture medium was replaced every 2 days during
the culture period for PGCs, and the cultured PGCs were passaged until 90% confluence. Several male PGCs were
proliferated to large numbers, and could be maintained for more than 100 days. The established PGC lines were
used for subsequent assays.
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