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Hyclone defined fetal bovine serum

Manufactured by GE Healthcare
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HyClone defined fetal bovine serum is a cell culture supplement derived from fetal bovine blood. It is designed to provide essential nutrients and growth factors to support the growth and proliferation of cultured cells.

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Lab products found in correlation

L glutamine, by Thermo Fisher Scientific (2 mentions) Dmem low glucose medium, by Thermo Fisher Scientific (1 mentions) M2 medium, by Merck Group (1 mentions) Penicillin streptomycin, by Merck Group (1 mentions) Noggin, by Merck Group (1 mentions) Acutase, by STEMCELL (1 mentions) Mtesr1, by Bio-Techne (1 mentions) Noggin, by R&D Systems (1 mentions) Chir99021, by Bio-Techne (1 mentions) Rpmi 1640 media, by Thermo Fisher Scientific (1 mentions) Sb431542, by Bio-Techne (1 mentions) Y 27632, by Bio-Techne (1 mentions) Activin a, by R&D Systems (1 mentions) Bone morphogenetic protein 4 (bmp4), by R&D Systems (1 mentions) Retinoic acid, by Merck Group (1 mentions) Wnt3a, by R&D Systems (1 mentions) Pen strep, by Thermo Fisher Scientific (1 mentions) Mycoalert mycoplasma detection kit, by Lonza (1 mentions) Mercaptoethanol, by Merck Group (1 mentions) Hg csf, by Merck Group (1 mentions)

4 protocols using hyclone defined fetal bovine serum

1

Imaging of LuVeLu Reporter Embryos

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Embryos expressing the LuVeLu reporter (Aulehla et al., 2008 (link)) were dissected in pre-warmed (at 37°C) M2 medium (Sigma) and cultured in low glucose DMEM medium (Gibco, 11054020), 10% of HyClone defined fetal bovine serum (GE Healthcare, #HYCLSH30070.03), 2 mM of L-glutamine (Gibco,#25030–024) and 1% penicillin-streptomycin (Sigma, #P0781). Embryos were cultured at 37°C in a 65% O2 and 15% CO2 environment (N2 balanced). Embryos were imaged on the Prairie two-photon system (laser tuned to 960 nm). At time = 0, a z-stack of 5 μm step-size was acquired at 1024 × 1024 pixel size, using the 20x objective. From t = 1 and onwards, we acquired z-stacks series of 1024 × 512 pixel images, spaced in depth at 10 μm, using a Nikon 16x LWD 0.8NA W objective. T-series were acquired every 8,5 min.
Dias A., Lozovska A., Wymeersch F.J., Nóvoa A., Binagui-Casas A., Sobral D., Martins G.G., Wilson V, & Mallo M. (2020). A Tgfbr1/Snai1-dependent developmental module at the core of vertebrate axial elongation. eLife, 9, e56615.
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2

Directed Differentiation of hPSCs to Anterior Foregut

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Confluent hPSC cultures were treated with Acutase (STEMCELL Technologies) to resuspend as single cells in mTeSR1 and Y-27632 (10 μM, Tocris) and plated on Matrigel. On the following day, differentiation into definitive endoderm was carried out as previously described (McCracken et al., 2014 (link)). Briefly, cells were treated with Activin A (100 ng mL-1, R&D systems, Minneapolis, MN) and BMP4 (50 ng mL-1, R&D systems) on the first day in RPMI 1640 media (Life Technologies). Cells in the following two days were treated with only Activin A (100 ng mL-1) in RPMI 1640 with increasing concentrations 0.2% and 2% of HyClone defined fetal bovine serum (dFBS, GE Healthcare Life Sciences).
For anterior foregut monolayer cultures, cells were treated for 3 days in Noggin (200 ng mL-1) in RPMI 1640 with 2% dFBS, with all-trans retinoic acid (2 μM, Sigma, St. Louis, MO) the 3rd day.
Alternately, for the generation of anterior foregut spheroids, from definitive endoderm, cells were treated with FGF4 (500 ng mL-1, R&D systems), Noggin (200 ng mL-1) for 3 days in RPMI 1640 with 2% dFBS. Additional factors were tested during this time (described in results), such as CHIR99021 (“chiron” or “chr”, 2 μM, Tocris), Wnt3a (500 ng mL-1, R&D systems), SB431542 (10 μM, Tocris), DEAB (10 μM, Sigma), and retinoic acid (2 μM).
Trisno S.L., Philo K.E., McCracken K.W., Cata E.M., RuizTorres S., Rankin S.A., Han L., Nasr T., Chatuvedi P., Rothenberg M.E., Mandegar M.A., Wells S.I., Zorn A.M, & Wells J.M. (2018). Esophageal organoids from human pluripotent stem cells delineate Sox2 functions during esophageal specification. Cell stem cell, 23(4), 501-515.e7.
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3

Prostate Cancer Cell Lines Cultivation

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The human cell lines LNCaP (androgen-sensitive human prostate adenocarcinoma cells) and 22RV1 (human prostate carcinoma epithelial cell line derived from a xenograft that was serially propagated in mice after castration-induced regression and relapse of the parental, androgen-dependent CWR22 xenograft) were obtained from ATCC and were cultured in RPMI-1640 medium. PC3 (metastatic prostate cancer cells isolated from bones, ATCC) was maintained in F-12K medium. All culture medium was supplemented with 10% HyClone Defined Fetal Bovine Serum (GE Healthcare) and 1% Pen/Strep (Invitrogen) unless specified. Cell cultures were tested every 6 months for cross-contamination using human STR profiling cell authentication service provided by ATCC. Mycoplasma contamination was tested using MycoAlert Mycoplasma Detection Kit (Lonza).
Shah K., Gagliano T., Garland L., O’Hanlon T., Bortolotti D., Gentili V., Rizzo R., Giamas G, & Dean M. (2020). Androgen receptor signaling regulates the transcriptome of prostate cancer cells by modulating global alternative splicing. Oncogene, 39(39), 6172-6189.
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4

Myeloid Lineage Differentiation Assay

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To test myeloid potential, single cells were cultured in Terasaki plates in 20 l of Iscove's Modified Dulbecco's Medium (IMDM) with l-glutamine (Gibco), 20% HyClone Defined Fetal Bovine Serum (SH30070.03, GE Healthcare), penicillin-streptomycin (Invitrogen), and 0.1 M -mercaptoethanol (Sigma-Aldrich), supplemented with hSCF (20 ng/ml), hFlt3L (20 ng/ml), hIl-3 (20 ng/ml), hIl-5 (50 ng/ml), hIl-6 (20 ng/ml), hGM-CSF (50 ng/ml), and hG-CSF (20 ng/ml). For bulk cultures, 50 to 150 cells were cultured in 400 l of the same culture medium in 48-well plates. For combined erythroid, Downloaded from https://www.science.org on September 09, 2024 megakaryocyte, and myeloid readout, single cells were cultured in round-bottom 96-well plates in 50 l of StemSpan (STEMCELL Technologies) with hSCF (20 ng/ml), hFlt3L (20 ng/ml), hIl-3 (20 ng/ml), hIl-5 (50 ng/ml), hIl-6 (20 ng/ml), hGM-CSF (50 ng/ml), hG-CSF (20 ng/ml), hLDL (40 g/ml; Sigma-Aldrich), erythropoietin (0.5 U/ml), and thrombopoietin (100 ng/ml). Cells were cultured at 37°C, 5% CO 2 . Cytokines, suppliers, and concentrations used are found in table S6. Cytospins were prepared with a Shandon Cytospin at 1000 rpm with low acceleration, followed by May-Grünwald-Giemsa stain (VWR).
Drissen R., Thongjuea S., Theilgaard-Mönch K, & Nerlov C. (2019). Identification of two distinct pathways of human myelopoiesis. Science immunology, 4(35).
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