Esc qualified fetal bovine serum
Manufactured by Thermo Fisher Scientific
Sourced in United States
ESC-qualified fetal bovine serum is a cell culture media supplement. It is derived from the blood of fetal bovine sources and is qualified for use in embryonic stem cell (ESC) research applications.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
β mercaptoethanol, by Merck Group (2 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (2 mentions)
Knockout dmem, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Dmem high glucose, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dmem f12, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Mitomycin c, by Thermo Fisher Scientific (1 mentions)
Pen strep antibiotics, by Thermo Fisher Scientific (1 mentions)
Knockout dmem, by Merck Group (1 mentions)
Mem non essential amino acid, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
L alanyl l glutamine, by Thermo Fisher Scientific (1 mentions)
Chir99021, by Bio-Techne (1 mentions)
Pd0325901, by Bio-Techne (1 mentions)
9 protocols using esc qualified fetal bovine serum
1
mESC and C2C12 Cell Culture
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Source cells. mESC line HBG3 (Hb9-GFP), a gift from H. Wichterle, Columbia University, NY, was kept in culture on a feeder layer of mouse embryonic fibroblasts (CF-1 MEF Feeder Cells, Applied StemCell) plated on 0.1% gelatin–coated dishes in undifferentiated medium consisting of EmbryoMax ES Dulbecco’s modified Eagle’s medium (DMEM) (Millipore Chemicon), 15% ESC-qualified fetal bovine serum (Invitrogen), 1% nucleosides (Millipore Chemicon), 1% nonessential amino acids (Millipore Chemicon), 1% penicillin-streptomycin (Invitrogen), 1% l -glutamine (Invitrogen), 0.1 mM β-mercaptoethanol (Sigma), and 0.1% leukemia inhibitory factor (EMD). Mouse myoblasts C2C12 (89 (link)) [American Type Culture Collection (ATCC)] were cultured below 70% confluency in growth medium consisting of high-glucose DMEM (ATCC), 10% fetal bovine serum (Invitrogen), and 1% penicillin-streptomycin. All cells were kept in incubators at 37°C and 5% CO2. None of the cells were used beyond a passage number 20.
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2
Sterilization and Culture of Embryonic Stem Cells
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We sterilized the nanofibrous scaffolds in 70%
ethanol for 2 hours after which they were washed
3 times with PBS for 20 minutes and subsequently
incubated with DMEM/F12 (Sigma-Aldrich, MO,
USA) for 24 hours before cell seeding. Mouse
embryonic fibroblast (MEF) cells were prepared
according to guidelines the Laboratory Animal
Ethical Commission of Tarbiat Modares University
and cultured in DMEM F12 supplemented
with 10% FBS (Sigma-Aldrich, MO, USA). After
reaching 70% confluency, the cells were chemically
inactivated with mitomycin C (10 μg/ml; Gibco-
Invitrogen, CA, USA) for 2 hours and washed
3 times with PBS to remove any remaining mitomycin
C. mESCs, obtained from the Stem Cell
Technology Research Center (17 (link)), were grown on
a gelatinized plate that contained one layer of inactive
feeder cells (MEFs) in ES culture medium
that included Knock-Out DMEM (Sigma-Aldrich,
MO, USA) which consisted of sodium pyruvate
supplemented with 20% ESC qualified fetal bovine
serum (Gibco-Invitrogen, CA, USA), 1000
IU/ml LIF (Gibco-Invitrogen, CA, USA), 1 mM
NEAA, 2 mM L-glutamine, 0.1 mM β-ME, and
100 μg/ml pen/strep antibiotics (Gibco-Invitrogen,
CA, USA). Cells were maintained in a humidified
CO2 incubator at 37˚C. Media was changed daily
until the appropriate confluency of ESC colonies
was attained.
ethanol for 2 hours after which they were washed
3 times with PBS for 20 minutes and subsequently
incubated with DMEM/F12 (Sigma-Aldrich, MO,
USA) for 24 hours before cell seeding. Mouse
embryonic fibroblast (MEF) cells were prepared
according to guidelines the Laboratory Animal
Ethical Commission of Tarbiat Modares University
and cultured in DMEM F12 supplemented
with 10% FBS (Sigma-Aldrich, MO, USA). After
reaching 70% confluency, the cells were chemically
inactivated with mitomycin C (10 μg/ml; Gibco-
Invitrogen, CA, USA) for 2 hours and washed
3 times with PBS to remove any remaining mitomycin
C. mESCs, obtained from the Stem Cell
Technology Research Center (17 (link)), were grown on
a gelatinized plate that contained one layer of inactive
feeder cells (MEFs) in ES culture medium
that included Knock-Out DMEM (Sigma-Aldrich,
MO, USA) which consisted of sodium pyruvate
supplemented with 20% ESC qualified fetal bovine
serum (Gibco-Invitrogen, CA, USA), 1000
IU/ml LIF (Gibco-Invitrogen, CA, USA), 1 mM
NEAA, 2 mM L-glutamine, 0.1 mM β-ME, and
100 μg/ml pen/strep antibiotics (Gibco-Invitrogen,
CA, USA). Cells were maintained in a humidified
CO2 incubator at 37˚C. Media was changed daily
until the appropriate confluency of ESC colonies
was attained.
3
Culturing Mouse Embryonic Stem Cells
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Mouse D3 embryonic stem cells (ESCs; ATCC™, Manassas, VA) were cultured as described previously [13 (link)]. Briefly, cells were cultured on gelatin-coated tissue culture plastic with medium that consisted of Dulbecco’s Modification of Eagles Medium, 15% ESC-qualified fetal bovine serum (Invitrogen, Carlsbad, CA), 2 mM L-glutamine, 0.1 mM non-essential amino acids, antibiotics, and 1000 U/ml leukemia inhibitory factor (LIF; EMD Millipore, Temecula, CA).
4
Mouse Embryonic Stem Cell Culture and Differentiation
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The experiments were performed using the mouse ESC line W4 (provided by the Rockefeller University Core Facility), the YPet-OCT4 and YPet-SOX2 ESC lines previously generated in our laboratory from the W4 cell line (Verneri et al., 2020 (link)) and the eGFP-NANOG cell line that was generated in this work. ESCs were maintained on 0.1% gelatin-coated dishes, passed every 3 days using trypsin-EDTA (Gibco) and grown at 37°C in a 5% CO2 (v/v) incubator. Cells were cultured in DMEM (Gibco) supplemented with 15% ESC qualified fetal bovine serum (Gibco), 100 mM MEM nonessential amino acids (Gibco), 2 mM l-alanyl-L-glutamine (Gibco), 0.5 mM 2-mercaptoethanol, 100 U/mL penicillin (Gibco), 100 mg/mL streptomycin (Gibco), leukemia inhibitory factor (LIF) and 2i [1 μM PD0325901 (Tocris) and 3 μM CHIR99021 (Tocris)].
To induce non-directed differentiation, ESCs were incubated in the absence of LIF and 2i during 4 h for induced early-S phase cells measurements and for 24 or 48 h for eGFP-NANOG stable cell line validation.
To induce non-directed differentiation, ESCs were incubated in the absence of LIF and 2i during 4 h for induced early-S phase cells measurements and for 24 or 48 h for eGFP-NANOG stable cell line validation.
5
Culturing Mouse Embryonic Stem Cells
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E14TG2a mouse ESCs (ATCC) were routinely cultured on gelatin-coated plates in DMEM (GIBCO) supplemented with 15% ESC-qualified fetal bovine serum (GIBCO), 10 mM 2-mercaptoethanol, 1mM nonessential amino acids, and 1,000 U/ml of LIF (Millipore). For cell maintenance, ESCs were cultured in the serum-free ESGRO medium (Millipore). 293T cells were maintained in DMEM with 10% fetal bovine serum (Biological Industries) at 37°C with 5% CO2.
6
Murine Embryonic Stem Cell Culture and Differentiation
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Commercially available mESCs (E14.1) were cultured on plates coated with 0.1% gelatin (Millipore, Billerica, MA, http://www. millipore.com/) in high-glucose Dulbecco's modified Eagle's medium (DMEM; HyClone, Waltham, MA, http://www.thermo. com.cn/) supplemented with 15% ESC-qualified fetal bovine serum (Gibco, Grand Island, NY, http://www.invitrogen.com), 1% penicillin and streptomycin (Gibco), 2 mM L-glutamine (Gibco), 0.1 mM nonessential amino acids (Gibco), 0.1 mM 2mercaptoethanol (Gibco), and 1,000 units/ml of LIF (Millipore). For differentiation without LIF, mESCs were cultivated in the culture medium without LIF. EBs were formed via aggregation in this culture medium. Additionally, 293FT cells were maintained in high-glucose DMEM containing 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin. For teratoma formation, nonobese diabetic mice with severe com-bined immunodeficiency underwent transplantation with 2 3 10 6 control or Pwp1 KD cells in their hind limbs. After 5 weeks, the subcutaneous teratomas were dissected and immunohistochemically stained with lineage-specific markers.
7
Derivation of Mouse Embryonic Stem Cells
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Mouse ESCs were derived by in vitro expansion of E3.5 blastocysts, which were obtained by breeding Upf2-heterozygous mice. Briefly, following the previously described protocol for isolation of mouse ESC from non-permissive lines, blastocysts were cultured on irradiated mouse embryonic feeders (MEFs) (Thermo Fisher Scientific, A34181) in a mixture of DMEM/F-12 and Neurobasal media with N2 and B27 supplements. The media was further supplemented with MEK (1 μM, PD325901) and GSK (3 μM, CHIR99021) inhibitors along with LIF (EMD Millipore, ESG1106) (2i-LIF) and 2% knockout serum replacement. Once blastocysts attached to the feeder layer, half media changes were carried out every 2 days. Cells were maintained in 2i-LIF media for 10-12 days. Large clusters of outgrown cells were then split and maintained either in 2i-LIF or in serum containing mouse ESC media containing KnockOut DMEM (Thermo Fisher Scientific, 10829018), 20% ESC-qualified fetal bovine serum (Thermo Fisher Scientific, 10439024), L-glutamine and supplemented with non-essential amino acids and 1000 U/ml of recombinant mouse Lif (EMD Millipore, ESG1106). Statistical analysis of comparisons between genotypes was conducted using two-tailed Student's t-tests.
8
Maintenance of Embryonic Stem Cells
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We maintained mouse embryonic stem cells (E14) in ESC Media containing: DMEM (Fisher Scientific) supplemented with 15 % ESC-qualified fetal bovine serum (Thermo Fisher Scientific), 1x non-essential amino acids (Thermo Fisher Scientific), 1x GlutaMAX (Thermo Fisher Scientific), 1x ESC-qualified nucleosides (EMD Millipore), 0.1 mM β-Mercaptoethanol (Sigma-Aldrich), and 103 units/ml ESGRO leukemia inhibitor factor (LIF) (EMD Millipore). N2a cells were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco, Invitrogen) supplemented with 10% fetal bovine serum (FBS) and penicillin-streptomycin. Cells were grown in incubator with 5% CO2 at 37C.
9
Feeder-free Culture of Embryonic Stem Cells
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ESCs were grown in feeder-free culture condition and incubated at 37°C under 3% O2 tension. ESC medium composed of KnockOut DMEM (ThermoFisher, 10829–018), 15% ESC qualified Fetal Bovine Serum (ThermoFisher, 16141–079), 2 mM L-glutamine, 1/500 home-made leukemia inhibitory factor (LIF), 0.1 mM non-essential amino acids, 0,1 mM 2-mercaptoethanol, 50 units/mL penicillin, 50 mg/mL streptomycin supplemented with the two inhibitors (2i); PD 0325901 (1 µM) and CHIR 99021 (3 µM) was used. MEFs were cultured in high glucose DMEM (Lonza, BE12-614F) supplemented with 10% North American FBS, 2 mM L-glutamine, 0.1 mM non-essential amino acids, 50 units/mL penicillin, 1 mM Sodium Pyruvate and 50 mg/mL streptomycin (MEF medium).
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