Es fbs

Manufactured by Thermo Fisher Scientific

Sourced in United States, Italy

ES-FBS is a serum product designed for cell culture applications. It is derived from fetal bovine sources and is processed to meet specific quality standards. The core function of ES-FBS is to provide a source of essential nutrients, growth factors, and other components necessary for the optimal growth and maintenance of a variety of cell types in vitro.

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Close spelling variants of this product

ES-qualified FBS (8 citations) ES Cell FBS (4 citations) ES-cell qualified FBS (4 citations) ES-grade FBS (3 citations) FBS-Williams’ Medium E (2 citations)

22 protocols using es fbs

Murine Embryonic Stem Cell Culture

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Murine ES v6.5 cells were purchased from Open Biosystems. mES cells were maintained in an undifferentiated, feeder-free state in leukemia inhibitory factor (LIF) supplemented medium (Knockout Dulbecco’s modified Eagle’s medium, Invitrogen, Carlsbad, CA) with 15% ES-FBS (Invitrogen), 0.1 mM β-mercaptoethanol, 2 mM L-glutamine (Invitrogen), 0.1 mM nonessential amino acids (Invitrogen), 1000 U/mL recombinant LIF (Chemicon, Temecula CA), and 2 mM HEPES (Invitrogen). Cells were cultured on gelatin-coated (0.1% gelatin in PBS, coated for 2h at 37°C) T-75 flasks at 37C, 5% CO₂ in a humidified incubator. Cells were passaged every 2–3 days to maintain an undifferentiated state.
For initial differentiation assays, mES cells were introduced to collagen IV flasks (BD Biosciences, San Jose, CA) and maintained in α-minimum essential medium (α-MEM) (Invitrogen) supplemented with 10% ES-FBS (Invitrogen), 0.1 mM β-mercaptoethanol, 2 mM L-glutamine (Invitrogen), 0.1 mM nonessential amino acids (Invitrogen), and 2 mM HEPES (Invitrogen). Media was refreshed daily. After 5 days, cells were isolated for magnetic-activated cell sorting (MACS).

Gluck J.M., Delman C., Chyu J., MacLellan W.R., Shemin R.J, & Heydarkhan-Hagvall S. (2014). Microenvironment influences vascular differentiation of murine cardiovascular progenitor cells. Journal of biomedical materials research. Part B, Applied biomaterials, 102(8), 1730-1739.

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Differentiation of Mouse ESCs into mLPCs

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Mouse ES-D3 [D3](ATCC CRL-1934) were propagated on growth-arrested mouse embryonic fibroblasts (CF-1 MEF, GlobalStem) in ES FBS medium consisting of DMEM supplemented with 15% ES FBS (Invitrogen), 2 mM L-glutamine, 1 mM MEM NEAA, 100 nM 2-mercaptoethanol, 1 × penicillin-streptomycin, and 10³ U/mL LIF (ESGRO, Millipore). Undifferentiated mouse ESCs were kept at 37 °C, 5% CO₂ and 90–95% humidity, with medium exchanged daily, and passaged every 3 days using trypsin-EDTA. Prior to differentiation, cells were cultured in 6-well tissue culture plates pre-coated with 0.1 mg/ml fibronectin for 24 hrs in ES FBS media. To initiate differentiation, cells were switched to endoderm differentiation media containing IMDM (Gibco) supplemented with 10% ES-FBS, 2 mM L-glutamine, 1 mM MEM NEAA, 100 nM 2-mercaptoethanol, 1 × penicillin-streptomycin, and 50 ng/ml Activin A (Invitrogen) for 4 days to generate definite endoderm (DE) cells. From day 5 through day 7 DE cells were cultured in the same differentiation media together with 50 ng/ml basic fibroblast growth factors (bFGF) to induce DE cells into mLPCs. Fresh media was exchanged every two days throughout the differentiation experiments.

Haque A., Gheibi P., Stybayeva G., Gao Y., Torok N, & Revzin A. (2016). Ductular reaction-on-a-chip: Microfluidic co-cultures to study stem cell fate selection during liver injury. Scientific Reports, 6, 36077.

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Generation of iPSCs from SOD1 Mice

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Neuronal progenitor cells (NPCs), expressing the motor neuron Hb9∷GFP reporter, obtained from SOD1^WT and SOD1^G93A mice were converted to iPSCs. As previously described, retrovirus encoding OCT3/4 and KLF4 were sufficient to generate iPSC clones (Hester et al., 2009 (link); Kim et al., 2008 (link)). Twenty viral particles per cell were needed to efficiently reprogram the cells. Cells were cultured in the presence of NPC media for four days followed by a change to mouse embryonic stem cell (mESC) media with DMEM (Millipore, Billerica, MA), supplemented with 18% ES FBS (Invitrogen), L-glutamine (2mM, Invitrogen), nonessential amino acids (1×, Millipore), antibiotic-antimycotic (Invitrogen), 2-mercaptoethanol (Sigma), and recombinant LIF (100U/ml, Millipore). iPSC clones were morphologically similar to mouse ESCs (HBG3 cells, Thomas Jessell, Columbia University) and were obtained within two weeks. A wide panel of markers was used to compare ESCs with the newly generated iPSC lines and found no significant difference for their expression between cell lines.

Israelson A., Ditsworth D., Sun S., Song S., Liang J., Hruska-Plochan M., McAlonis-Downes M., Abu-Hamad S., Zoltsman G., Shani T., Maldonado M., Bui A., Navarro M., Zhou H., Marsala M., Kaspar B.K., Da Cruz S, & Cleveland D.W. (2015). Macrophage Migration Inhibitory Factor (MIF) as a Chaperone Inhibiting Accumulation of Misfolded SOD1. Neuron, 86(1), 218-232.

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Generating iPSCs from SOD1 Mice

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NPCs, expressing the MN Hb9::GFP reporter, obtained from wild-type and SOD1^G93A mice were converted to iPSCs. As previously described, retrovirus encoding OCT3/4 and KLF4 were sufficient to generate iPSC clones^{63 (link),64 (link)}. 20 viral particles per cell were needed to efficiently reprogram the cells. Cells were cultured in the presence of NPC media for four days followed by a change to mouse embryonic stem cell (mESC) media with DMEM (Millipore, Billerica, MA), supplemented with 18% ES FBS (Invitrogen), L-glutamine (2mM, Invitrogen), nonessential amino acids (1x, Millipore), antibiotic-antimycotic (Invitrogen), 2-mercaptoethanol (Sigma), and recombinant LIF (100U/ml, Millipore). iPSC clones were morphologically similar to mouse ESCs (HBG3 cells, Thomas Jessell, Columbia University) and were obtained within two weeks. A wide panel of markers was used to compare ESCs with the newly generated iPSC lines.

Song S., Miranda C.J., Braun L., Meyer K., Frakes A.E., Ferraiuolo L., Likhite S., Bevan A.K., Foust K.D., McConnell M.J., Walker C.M, & Kaspar B.K. (2016). MHC class I protects motor neurons from astrocyte-induced toxicity in amyotrophic lateral sclerosis (ALS). Nature medicine, 22(4), 397-403.

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Culturing and Differentiating Trophoblast Stem Cells

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TSCs isolated from C57BL/6 laboratory mice were cultured in PMEF-conditioned medium (RPMI) supplemented with 10% ES-FBS (Invitrogen), 1% Antibiotic-Antimycotic (Invitrogen), 25 ng/ml FGF4 (Sigma), 1 μg/ml Heparin (Sigma), 0.1 mM 2-Mercaptoethanol (Sigma), and 1 mM sodium pyruvate (Sigma) at 37 °C in a humidified chamber with 5% CO₂ as described elsewhere [15 (link), 22 ]. The differentiation of TSCs into polyploid TGCs was induced by growing the cells in RPMI medium supplemented only with 10% FBS and 1% Antibiotic-Antimycotic. For the best differentiation, TSCs were kept in differentiation medium for 4 days. For subculturing, TSCs were detached from the plates with 0.05% trypsin (Sigma-Aldrich) for 5–10 min. trypsin was inactivated by addition of culture medium to the flasks; cells were centrifuged and resuspended in culture medium and replated at the desired density.

Ullah R., Naz A., Akram H.S., Ullah Z., Tariq M., Mithani A, & Faisal A. (2020). Transcriptomic analysis reveals differential gene expression, alternative splicing, and novel exons during mouse trophoblast stem cell differentiation. Stem Cell Research & Therapy, 11, 342.

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Generating iPSCs from SOD1 Mice

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Expansion and Drug Screening of Umbilical Cord Hematopoietic Stem Cells

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Human umbilical cord blood was obtained after caesarean birth. Informed consent was obtained from the parents and the study was approved by the local ethics committee. CD34⁺ cells were enriched by MACS-technology (Miltenyi), cell purity was generally more than 90%. Purified cells were frozen in CryoStor CS10 (Stem Cell Technologies), stored in liquid nitrogen and used at later time points. Thawed cells were cultured in serum-free medium supplemented with 10% ES-FBS (Invitrogen), human TPO (50 ng/mL, Immunotools), FLT3L, SCF, IL3 (100 ng/mL each, Immunotools), human EGF (hEGF) (20 or 200 ng/mL Immunotools), PGE2 (10, 25 or 50 μM, Sigma-Aldrich) and/or cytotoxic drugs (etoposide, taxol, tunicamycin; Sigma-Aldrich). Alternatively, thawed cells were used for xenotransplantation.

Demmerath E.M., Bohler S., Kunze M, & Erlacher M. (2019). In vitro and in vivo evaluation of possible pro-survival activities of PGE2, EGF, TPO and FLT3L on human hematopoiesis. Haematologica, 104(4), 669-677.

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Isolation and Culture of Pediatric Adipose-Derived Stem Cells

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Abdominal adipose tissue was collected from consenting patients under ethical approval from the Camden and Islington Community Local Research Ethics Committee (London, UK). ADSCs were isolated from lipoaspirates of pediatric patients as previously described17 (link). Isolated ADSCs were cultured in high glucose Dulbecco’s modified Eagle’s medium (DMEM; Life Technologies) containing GlutaMAX™ and supplemented with 10% embryonic stem cell-qualified fetal bovine serum (ES-FBS; Invitrogen, Carlsbad, CA) and 1% penicillin/streptomycin.

Vagaska B., New S.E., Alvarez-Gonzalez C., D’Acquisto F., Gomez S.G., Bulstrode N.W., Madrigal A, & Ferretti P. (2016). MHC-class-II are expressed in a subpopulation of human neural stem cells in vitro in an IFNγ–independent fashion and during development. Scientific Reports, 6, 24251.

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Stepwise Differentiation of Stem Cells

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Prior to differentiation, cells were cultured inside microfluidic chambers or in 12-well tissue culture plates precoated with 0.1% gelatin for 24 hours in ES FBS media. To initiate differentiation, cells were switched to differentiation media containing DMEM supplemented with 15% ES FBS (Invitrogen), 2 mM L-glu- tamine, 1 mM MEM NEAA, 100 nM 2-mercaptoethanol, and 13penicillin-streptomycin, and 1 mM Jak1 inhibitor (J1I; Calbiochem, Millipore, Billerica, MA, http://www.emdmillipore.com) for 5 days. Five micromolar of dorsomorphin (an inhibitor of BMP-mediated Smad1/5/8 phosphorylation, Stemgent) was utilized where indicated. From day 5 through day 10 cells were cultured in 1% ES FBS, 2 mM L-glutamine, 1 mM MEM NEAA, 100 nM 2-mercaptoethanol, and 13penicillin-streptomycin. Fresh media was exchanged daily throughout all experiments.

Guild J., Haque A., Gheibi P., Gao Y., Son K.J., Foster E., Dumont S, & Revzin A. (2016). Embryonic Stem Cells Cultured in Microfluidic Chambers Take Control of Their Fate by Producing Endogenous Signals Including LIF. Stem cells (Dayton, Ohio), 34(6).

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Culturing Mutant Mouse Embryonic Stem Cells

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10 cm plates were coated with 0.1% gelatin generally overnight. Cells (E14 WT, EED−/−^{41 (link)} and RING1a−/−RING1b^fl/fl42 mESC) were thawed and resuspended in 10 ml of medium (serum + LIF). Standard serum + LIF medium contained 500 mL Knockout DMEM (Gibco 10829-018), 95 mL of filtered ES FBS (Gibco 16141-061), 5.9 mL of antibiotics (Hyclone SV30079.01), 5.9 mL Glutamax (Gibco 35050-038), 5.9 mL MEM NEAA (Gibco 11140-035), 4.7 mL titrated LIF (Miltenyi Biotec 130-095-777), and 1.3 mL Beta-mercaptoethanol 55 mM (Gibco 21985-023). Cells were split once (1/3 or 1/4) every two days. Ring1a−/−Ring1b^fl/fl mESC were treated with Tamoxifen (OHT; 1 mM) for 72 h right after one passage and RING1B loss was confirmed by PCR genotyping and Western Blot.

Crispatzu G., Rehimi R., Pachano T., Bleckwehl T., Cruz-Molina S., Xiao C., Mahabir E., Bazzi H, & Rada-Iglesias A. (2021). The chromatin, topological and regulatory properties of pluripotency-associated poised enhancers are conserved in vivo. Nature Communications, 12, 4344.

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