Fetal calf serum (fcs)

Manufactured by GE Healthcare

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The FCS is a lab equipment product by GE Healthcare that is designed to measure and analyze fluorescence signals in samples. It provides accurate quantification and characterization of fluorescent molecules, particles, and cells.

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Calf serum (10 citations)

344 protocols using fetal calf serum (fcs)

Cell Culture Protocols for X-Inactivation Studies

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Mouse-human somatic cell hybrid cell lines, t75-2maz 34-1a (containing a human Xi) and t60-12 (containing a human Xa) [52 (link)] were cultured at 37°C in alpha Minimum Essential Medium (MEM) supplemented with 7.5% fetal calf serum (PAA Laboratories Inc), 1% penicillin/streptomycin (Life Technologies) and 1% L-glutamine (Life Technologies). The GM11200 and GM7057 male lymphoblast cell lines and GM11201, and GM7350 female lymphoblast cells lines (Coriell cell repository) were maintained in Roswell Park Memorial Institute (RPMI) medium supplemented with 15% fetal calf serum (PAA Laboratories Inc), 1% penicillin/streptomycin (Life Technologies) and 1% L-glutamine (Life Technologies). The IMR90 female fibroblast cell line was cultured with 10% fetal calf serum (PAA Laboratories Inc) and 1% L-glutamine (Life Technologies). The HT1080XISTi transgenic cell line containing a DOX-inducible XIST was cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal calf serum (PAA Laboratories Inc), 1% penicillin/streptomycin (Life Technologies) and 1% L-glutamine (Life Technologies). CA1S cells were cultured as described [53 (link)] on Matrigel (BD Biosciences) coated 6-well plates in mTeSR1 basal medium (STEMCELL) supplemented with mTeSR1 5x supplement (STEMCELL) and passaged using Accutase (STEMCELL). All lines were cultured at 37°C.

Chapman A.G., Cotton A.M., Kelsey A.D, & Brown C.J. (2014). Differentially methylated CpG island within human XIST mediates alternative P2 transcription and YY1 binding. BMC Genetics, 15, 89.

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Cell Culture Conditions for Microfluidic Assays

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The cat brain Moloney sarcoma virustransformed PG-4 cell line (European Collection of Cell Cultures, 94102703) and the normal mouse tail fibroblast M. dunni (Clone III8C) cell line (European Collection of Cell Culture, 94101211) were cultured at 37 °C in 5% CO 2 humidified atmosphere (incubator; HeraCell). Cell expansion took place in 25 cm 2 cell culture flaks (PAA Laboratories), where after reaching 70 to 80% confluence both cell lines were split at a ratio of 1 : 6 using 0.25% trypsin-EDTA (trypsinethylenediaminetetraacetic acid, fisher scientific) at 37 °C for 3 min for enzymatic cell detachment. The culture medium, McCoys (PAA Laboratories), was supplemented with 1% stabile L-glutamine (PAA Laboratories) and 5% FCS (PAA Laboratories) for the M. dunni cell line as well as 10% FCS for die PG-4 cell line. For on-chip experiments cells were transferred into a 6-well plate 24 h prior to their seeding into the microfluidic device at a density of approximately 70% to ensure comparability of the cellular state between experiments. The final cell culture medium was prepared with 10% FCS and complemented with 15 mM HEPES puffer (PAA Laboratories) for on-chip experiments.

Purtscher M., Rothbauer M., Kratz S.R.A., Bailey A., Lieberzeit P, & Ertl P. (2021). A microfluidic impedance-based extended infectivity assay: combining retroviral amplification and cytopathic effect monitoring on a single lab-on-a-chip platform. Lab on a chip, 21(7).

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Trout Spleen Cell Isolation

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Rainbow trout were killed by benzocaine (Sigma-Aldrich) overdose and the spleen collected. Single cell suspensions were obtained as previously reported^{18 (link),24 (link)} using 100 µm nylon cell strainers (BD Biosciences) and Leibovitz medium (L-15, Invitrogen) supplemented with 100 I.U./ml penicillin and 100 µg/ml streptomycin (P/S, Life Technologies), 10 units/ml heparin (Sigma) and 5% foetal calf serum (FCS, Life Technologies). Cell suspensions were placed onto 30/51% discontinuous Percoll (GE Healthcare) density gradients and centrifuged at 500 × g for 30 min at 4 ºC. The interface cells were washed twice in L-15 with 2% FCS and cells were resuspended in L-15 with 5% FCS. The viable cell concentration was determined by Trypan blue (Sigma-Aldrich) exclusion, adjusting the concentration to 2 × 10⁶ cells/ml.

Soleto I., Morel E., Muñoz-Atienza E., Díaz-Rosales P, & Tafalla C. (2020). Aeromonas salmonicida activates rainbow trout IgM+ B cells signalling through Toll like receptors. Scientific Reports, 10, 16810.

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Cell Culture Conditions for Neuroblastoma, Liver, and Lung Cancer

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The human neuroblastoma cell line SH-SY5Y wild-type was cultivated in Dulbecco’s modified Eagle’s medium (DMEM) with 10% fetal calf serum (FCS, GE Healthcare Life Sciences, Chalfont St. Giles, United Kingdom) and 1% non-essential amino acid solution at 37 °C and 5% CO₂. For cultivation of the liver hepatocellular carcinoma cell line HEPG2 wildtype, 1% penicillin/streptomycin was additionally added to the medium. The lung cancer cell line Calu-3 wild-type was cultivated in Dulbecco’s modified Eagle’s medium–F12 (DMEM F12) with 10% fetal calf serum (FCS, GE Healthcare Life Sciences, Chalfont St. Giles, United Kingdom) at 37 °C and 5% CO₂. All cells were seeded out on six-well plates uniformly and cultivated to a confluency of 90%–100% before starting treatment with methylxanthines.

Janitschke D., Lauer A.A., Bachmann C.M., Seyfried M., Grimm H.S., Hartmann T, & Grimm M.O. (2020). Unique Role of Caffeine Compared to Other Methylxanthines (Theobromine, Theophylline, Pentoxifylline, Propentofylline) in Regulation of AD Relevant Genes in Neuroblastoma SH-SY5Y Wild Type Cells. International Journal of Molecular Sciences, 21(23), 9015.

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Protocol for Culturing HepG2 and Porcine Hepatocytes

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Human liver hepatoma cells HepG2 were grown in Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen, Karlsruhe, Germany) supplemented with 10 % FCS (GE Healthcare), 100 μg/mL of streptomycin, 100 IU/mL of penicillin (Invitrogen), 2 mM L-glutamine and 1 % nonessential amino acids (Invitrogen) (DMEM complete) at 37 °C in a 5 % (vol/vol) CO₂ incubator. Primary porcine hepatocytes (PPHs) were isolated from liver specimens by perfusion technique as previously described (Fráguas-Eggenschwiler et al., 2021 (link)). Hepatocytes were plated at a density of 3 × 10⁵ on collagen-precoated 24-well plates, and kept in hepatocyte culture medium (William ´s medium E; Invitrogen) supplemented with 5 % FCS (GE Healthcare), 100 μg/mL of streptomycin, 100 IU/mL of penicillin (Invitrogen), 1 % GlutaMAX (Invitrogen), 2 % nonessential amino acids (Invitrogen), 10 mM HEPES buffer (Invitrogen), 2 % Dimethylsulfoxid HybriMax (Merck), 5 μg/mL insulin (Sigma-Aldrich), 5.41 μM hydrocortisone, 5.5 ng/mL epidermal growth factor (EGF) (Med Chem Express).

Praditya D.F., Klöhn M., Brüggemann Y., Brown L.E., Porco JA J.r., Zhang W., Kinast V., Kirschning A., Vondran F.W., Todt D, & Steinmann E. (2022). Identification of structurally re-engineered rocaglates as inhibitors against Hepatitis E virus replication. Antiviral research, 204, 105359.

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Cell Culture and Transfection Protocols

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HEK293T cells were grown in DMEM with 10 % (v/v) FCS (GE Healthcare), 2 mM L-glutamine (GE Healthcare) and 1 % (v/v) Pen Strep (GE Healthcare) at 37°C and 5 % CO₂. SEM cells were cultured under the same conditions with RPMI 1640 supplemented with 10 % (v/v) FCS and 1 % (v/v) Pen Strep (GE Healthcare). Transient and stable transfections of HEK293T cells were performed with polyethylenimine (PEI). The transfections of SEM cells were performed with the AMAXA^® Nucleofector^® (Kit R, program T-016, Lonza AG).

Kühn A., Löscher D, & Marschalek R. (2016). The IRX1/HOXA connection: insights into a novel t(4;11)- specific cancer mechanism. Oncotarget, 7(23), 35341-35352.

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Culturing HPV-Negative Cervical Cancer Cells

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The HPV-negative human cervical cancer cell line C33A was cultured in DMEM High Glucose (PAA Laboratories, Cölbe, Germany) supplemented with 10% FCS (PAA Laboratories, Cölbe, Germany) and 1% gentamicin solution (PAA Laboratories, Cölbe, Germany). The C33A cell line was kindly provided by Frank Stubenrauch, PhD (UKT, University of Tübingen; [24] (link)). Authenticity of C33A cells was verified by the DSMZ GmbH (Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany). C33A-RFP cells were cultured under same conditions except for adding 5 µg/mL blasticidin. Human epithelial kidney cells (293FT) were obtained from Invitrogen GmbH (Karlsruhe, Germany) and cultured in RPMI 1640 supplemented with 10% FCS and 2 mM L-glutamine (PAA Laboratories, Cölbe, Germany).

Donat U., Rother J., Schäfer S., Hess M., Härtl B., Kober C., Langbein-Laugwitz J., Stritzker J., Chen N.G., Aguilar R.J., Weibel S, & Szalay A.A. (2014). Characterization of Metastasis Formation and Virotherapy in the Human C33A Cervical Cancer Model. PLoS ONE, 9(6), e98533.

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Cell Culture Conditions for In Vitro Studies

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HepaRG™ cells were purchased from Life Technologies (Waltham, USA) and they were plated and maintained in Williams’ medium E supplemented with GlutaMAX and HepaRG Thaw, Plate & General Purpose Medium Supplement (Thaw, Plate, & General Purpose Working Medium) (all purchased from Life Technologies). Vincristine-resistant (VCR-R) CEM cells were a kind gift from Prof. Maria Kavallaris (University of New South Wales, Australia) and cultivated with RPMI 1640 (PAN Biotech, Aidenbach, Germany), supplemented with 10% fetal calf serum (FCS) (PAA Laboratories, Toronto, Canada). HepG2 cells were obtained from German Research Centre of Biological Material (DSMZ) and cultivated with DMEM (PAN Biotech), supplemented with 10% fetal calf serum (FCS) (PAA Laboratories). Human umbilical vein endothelial cells (HUVECs) were purchased from Promocell and cultivated with ECGM Kit enhanced (PELO Biotech, Planegg, Germany) supplemented with 10% (FCS) (PAA Laboratories) and 1% penicillin/streptomycin/amphotericin B (all purchased from PAN Biotech). HUVECs were cultured for a maximum of six passages. All cells were cultured at 37 °C with 5% CO₂ with constant humidity are proven to be mycoplasma-free on a quarterly basis.

Schütz R., Müller M., Geisslinger F., Vollmar A., Bartel K, & Bracher F. (2020). Synthesis, biological evaluation and toxicity of novel tetrandrine analogues. European Journal of Medicinal Chemistry, 207, 112810.

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Breast Cancer Cell Culture Protocols

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The cell lines BT20, BT474, MDA-MB-468, T47D, MCF-7, GI101 and MDA-MB-231 (breast adenocarcinoma cells) and MTSV1.7 (breast epithelial cells) were cultured in DMEM (Invitrogen, Karlsruhe, Germany) supplemented with 10% FCS (fetal calf serum; PAA Laboratories, Cölbe, Germany), 2 mM L-glutamin (Invitrogen) and under standard conditions (37 °C, 10% CO₂, humidified atmosphere). The micrometastatic cell line BCM1 was cultured at 37 °C, 5% CO₂ and 10% O₂ in RPMI (Invitrogen, Karlsruhe, Germany) supplemented with 10% FCS (PAA Laboratories), 2 mM L-glutamin (Invitrogen), 10 mg ml⁻¹ Insulin-Transferrin-Selenium-A (Invitrogen), 50 ng ml⁻¹ recombinant human epidermal growth factor, and 10 ng ml⁻¹ human basic fibroblast growth factor (both from Macs Miltenyi Biotec, Bergisch-Gladbach, Germany). Cell viability was determined by trypan blue staining.

Joosse S.A., Müller V., Steinbach B., Pantel K, & Schwarzenbach H. (2014). Circulating cell-free cancer-testis MAGE-A RNA, BORIS RNA, let-7b and miR-202 in the blood of patients with breast cancer and benign breast diseases. British Journal of Cancer, 111(5), 909-917.

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Cell Culture Protocols for Cancer Research

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HeLa cells were purchased from German Research Centre of Biological Material (DSMZ) and cultured in DMEM (PAN Biotech) supplemented with 10% FCS (PAA Laboratories). VCR-R CEM cells were a kind gift from Prof. Maria Kavallaris (University of New South Wales) [58 (link)] and cultured in RPMI-1640 (PAN Biotech) supplemented with 10% FCS (PAA Laboratories). Cells were cultured at 37 °C with high humidity and 5% CO₂.

Keller M., Sauvageot-Witzku K., Geisslinger F., Urban N., Schaefer M., Bartel K, & Bracher F. (2021). The ethoxycarbonyl group as both activating and protective group in N-acyl-Pictet–Spengler reactions using methoxystyrenes. A short approach to racemic 1-benzyltetrahydroisoquinoline alkaloids. Beilstein Journal of Organic Chemistry, 17, 2716-2725.

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