Medium 199

Manufactured by Corning

Sourced in United States

Medium 199 is a cell culture medium formulated for the growth and maintenance of a variety of cell types. It is a standard product used in many laboratory applications.

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Lab products found in correlation

Zeocin, by Thermo Fisher Scientific (4 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions) Mcdb 105, by Corning (3 mentions) Leibovitz l 15, by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin liquid, by Solarbio (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Medium 199, by Thermo Fisher Scientific (2 mentions) Penicillin and streptomycin, by Corning (2 mentions) Gentamicin sulfate, by Corning (2 mentions) Hyclone 2 mm l glutamine, by GE Healthcare (2 mentions) Dmem media, by Lonza (2 mentions) Mycoalert mycoplasma detection kit, by Lonza (2 mentions) Leibovitz l 15 and mccoy 5a lm media, by Thermo Fisher Scientific (2 mentions) Collagenase type 2, by Merck Group (2 mentions) Ham s f 10, by Corning (1 mentions) Skov3 cells, by American Type Culture Collection (1 mentions) Doxorubicin, by Selleck Chemicals (1 mentions) Cisplatin, by Selleck Chemicals (1 mentions) Zosuquidar, by Selleck Chemicals (1 mentions) Paclitaxel, by Selleck Chemicals (1 mentions)

24 protocols using medium 199

Chick Cell Isolation and Maintenance

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Chick kidney cells (CKCs) were prepared from 2–4 weeks-old specific-pathogen-free (SPF) chickens, obtained from the University of Illinois at Urbana-Champaign (UIUC) Poultry Farm, following standard methods [27 ] and seeded in growth medium consisting of Medium 199 (Cellgro, Corning, NY, USA) supplemented with 10% tryptose-phosphate broth (TPB), 0.63% NaHCO₃ solution, and antibiotics (100 U/mL penicillin and 100 µg/mL streptomycin) and 4% fetal bovine serum (FBS). Confluent CKCs were maintained in F10.199 medium consisting of a 1:1 mixture of Ham’s F10 (Cellgro) and Medium 199 supplemented with 7.5% TPB, 0.63% NaHCO₃, 0.2% FBS, and antibiotics.
Chicken embryo cells (CECs) were prepared from 10–11-day-old SPF embryos obtained from the UIUC Poultry Farm following standard methods [27 ]. Briefly, primary CECs were seeded in growth medium consisting of Medium 199 supplemented with 10% TPB, 0.63% NaHCO₃ solution, antibiotics, and 4% FBS. Confluent CECs were maintained in Medium 199 supplemented with 7.5% TPB, 0.63% NaHCO₃, 0.2% FBS, and antibiotics.
The chicken DF-1-Cre fibroblast cell line [28 (link)] was cultivated in a 1:1 mixture of Leibovitz L-15 and McCoy 5A (LM) media (Gibco, Gaithersburg, MD, USA) supplemented with 10% FBS and antibiotics, and maintained in 50 µg/mL Zeocin (Invitrogen, Carlsbad, CA, USA).
All cells were maintained at 38 °C in a humidified atmosphere of 5% CO₂.

Vega-Rodriguez W., Ponnuraj N., Garcia M, & Jarosinski K.W. (2021). The Requirement of Glycoprotein C for Interindividual Spread Is Functionally Conserved within the Alphaherpesvirus Genus (Mardivirus), but Not the Host (Gallid). Viruses, 13(8), 1419.

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Grass Carp Cell Culture Protocol

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Grass carp were obtained from Nanchang Shenlong Fisheries Development (Jiangxi, China) and domesticated in our laboratory. The breeding conditions were described in our previous study (35 (link)). CIK (C. idella kidney) cells and CO (C. idella Overy) cells from Professor Pin Nie (Institute of Hydrobiology, Chinese Academy of Sciences) were applied in the experiments. Both CIK cells and CO cells needed Medium 199 (CORNING, USA), to which was added 10% fetal bovine serum (FBS) and 0.6% penicillin–streptomycin liquid (Beijing Solarbio Science & Technology, China); the mixture was then cultured at 28°C in a cell incubator. In addition, CO cells require 5% CO₂.

Jiang Z., Sun Z., Hu J., Li D., Xu X., Li M., Feng Z., Zeng S., Mao H, & Hu C. (2022). Grass Carp Mex3A Promotes Ubiquitination and Degradation of RIG-I to Inhibit Innate Immune Response. Frontiers in Immunology, 13, 909315.

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Establishing Cisplatin-Resistant SKOV3 Cell Line

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SKOV3 cells were purchased from American Type Culture Collection (ATCC, Manassas VA). SKOV3 was grown in a 50:50 mixture of MCDB (Sigma Aldrich) and Medium 199 (Corning) media supplemented with 10% fetal bovine serum (FBS) and 100U penicillin/mL-100μg/mL streptomycin. To induce Pgp expression, the native SKOV3 cell line was exposed to increasing concentrations of Cisplatin (up to 1μM) by serial passage over 5–6 weeks to create the SKOV3cis cell line. Cisplatin, paclitaxel, doxorubicin, and zosuquidar were all purchased from SelleckChem (Houston, TX) and stocks prepared in dimethylformamide (Cisplatin) or dimethyl sulfoxide (DMSO). Experiments with zosuquidar (5 μM) included a 30 min pre-incubation before adding doxorubicin.

Narayanan G., Merrill D., An R., Nolte D.D, & Turek J.J. (2019). Intracellular Doppler Spectroscopy Detects Altered Drug Response in SKOV3 Tumor Spheroids with Silenced or Inhibited P-glycoprotein. Biochemical and biophysical research communications, 514(4), 1154-1159.

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Isolation and Culture of Porcine Islets

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One- to two-week old pre-weaned Yorkshire pig islets were isolated under procedures approved by the University of California Irvine, Institutional Animal Care and Use Committee. Pancreata were procured and stored in cold HBSS (<1 hour). Pancreata were minced and digested with Collagenase Type V (Sigma-Aldrich) in a 37°C shaking water bath for 15 minutes. The enzymatic digestion was quenched with HBSS supplemented with 1% porcine serum (Gibco-Thermo Fisher Scientific) and digested tissues were filtered through a 500 μm metal mesh. Islets were then cultured in maturation media made up of Ham’s F-12 medium (Corning), HEPES (Sigma-Aldrich), L-glutathione (Sigma-Aldrich), nicotinamide (Sigma-Aldrich), ITS+3 (Sigma-Aldrich), gentamycin sulfate (Corning), Trolox (Sigma-Aldrich), heparin (Sagent Pharmaceuticals), Pefabloc (Santa Cruz Biotechnology), L-glutamine(Alfa Aesar), medium 199 (Corning) and 10% porcine serum.

Medina J.D., Alexander M., Hunckler M.D., Fernández-Yagüe M.A., Coronel M.M., Smink A.M., Lakey J.R., de Vos P, & García A.J. (2020). Functionalization of alginate with extracellular matrix peptides enhances viability and function of encapsulated porcine islets. Advanced healthcare materials, 9(9), e2000102.

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Investigating ET-1 Signaling in Brain Endothelial Cells

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Experiments were performed in male-derived human brain microvascular endothelial cell line HBEC-5i (American Type Culture Collection-ATCC, CRL 3245). Cells were cultured in 75 cm² culture flasks that were coated with 0.2% w/v gelatin (porcine Type A; Sigma-Aldrich) before cell seeding. A 1:1 ratio of endothelial growth media (VEC Technologies, Rensselaer, NY, USA) and Medium 199 (Corning, Manassas, VA, USA) was used for cell culture. The VEC media includes serum and antibiotics, while 10% FBS and 1% penicillin-streptomycin were added to the M199. Cells were incubated with ET-1 (1μM) in the presence/absence of ETA receptor antagonist BQ123 (20 μM; cells were treated 30 minutes before ET-1 treatment) for 16 hours. Experimental design is depicted in Figure 1. Dose of ET-1 and BQ123 were selected based on our previous reports (Abdul et al. 2020a (link); Abdul et al. 2020b (link)). BQ123 was dissolved in distilled water and further diluted with working media. Thus, the control group did not receive vehicle treatment. The cell lysate was collected for Western blot analysis and RT-qPCR.

Smith H.O., Tabiti K., Schaffner G., Soldati D., Albrecht U, & Birnstiel M.L. (1991). Two-step affinity purification of U7 small nuclear ribonucleoprotein particles using complementary biotinylated 2'-O-methyl oligoribonucleotides.. Proceedings of the National Academy of Sciences of the United States of America, 88(21), 9784-9788.

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Culturing Human Cardiac Progenitor and Endothelial Cells

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All cell lines were preserved in a humidified incubator at 37°C, 5% CO₂ and atmospheric O₂. Human cardiac progenitor cells (hCPCs)^{[60 (link)]} and human coronary artery endothelial cells (HCAECs) were used between passages 17-23 and 7-14, respectively. hCPCs were cultured as previously described.^{[63 (link)]} Briefly, cells were cultured in 0.1% porcine gelatin (Sigma-Aldrich) coated flasks in growth media consisting of 10% fetal bovine serum (Thermo Fisher Scientific), 22% EBM2 (Lonza) complemented with EGM2 single quotes (Lonza) in Medium 199 (Corning), 1X non-essential amino acids (Lonza), and 1X penicillin-streptomycin (Life Technologies). HCAECs were grown in MesoEndo cell growth media (Cell Applications).

Hernandez M.J., Gaetani R., Pieters V.M., Ng N.W., Chang A.E., Martin T.R., van Ingen E., Mol E.A., Sluijter J.P, & Christman K.L. (2018). Decellularized Extracellular Matrix Hydrogels as a Delivery Platform for MicroRNA and Extracellular Vesicle Therapeutics. Advanced therapeutics, 1(3), 1800032.

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Culturing Ovarian and Fallopian Epithelial Cell Lines

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The SV40 transformed primary normal ovarian epithelial cell line (OSE tsT/hTERT, henceforth OSE) (18 (link)) was a kind gift from Dr. V. Shridhar, Mayo Clinic, Rochester, MN, USA. The SV40 transformed primary normal fallopian tube epithelial cells (henceforth FTE188) (19 (link)) were kindly provided by Dr. Jinsong Liu, MD Anderson Cancer Center, Houston, TX, USA. The CP20 cell line was a kind gift from Dr. Anil K. Sood, MD Anderson Cancer Center, Houston, TX, and was authenticated by the STR profiling facility at MD Anderson Cancer Center. The OV90, and OVCAR4 cell lines were purchased from ATCC and NCI respectively and OVSAHO, TYKNU, KURAMOCHI, OVKATE were purchased from JCRB Cell Bank (Japanese Collection of Research Bioresources Cell Bank). OSE cells were routinely cultured in 1:1 MCDB 105 and Medium 199 (Corning, Corning, NY, USA) + 10% FBS (Gibco, Grand Island, NY); FTE188 cells were cultured in 1:1 MCDB 105 and Medium 199 + 10% FBS + 0.01ug/ml EGF; CP20, OV90 and OVCAR4 were routinely cultured in RPMI + 10% FBS. All the cells were cultured with 1× penicillin-streptomycin (Gibco, Grand Island, NY) in a 5% CO₂ humidified atmosphere and tested for mycoplasma contamination prior to any experiment.

Dwivedi S.K., Khader S., Dey A., Mustafi S.B., Xiong X., Bhattacharya U., Neizer-Ashun F., Rao G., Wang Y., Ivan C., Yang D., Dudley J.T., Xu C., Wren J.D., Mukherjee P, & Bhattacharya R. (2019). KRCC1: a potential therapeutic target in ovarian cancer. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 34(2), 2287-2300.

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Immortalized HUVECs for Long-Term Studies

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Primary HUVECs freshly isolated from two different vascular beds were cultured in Medium 199 (Corning, Corning, NY, USA), supplemented with l-glutamine (Corning, Corning, NY, USA), antibiotic antimycotic (Corning, Corning NY, USA), HEPES buffer (Corning, Corning, NY, USA), heparin (Sigma Aldrich, St. Louis, MO, USA), and 20% fetal bovine serum (Hyclone, Logan, UT, USA). The HUVECs were transfected with the E4ORF1 of the Ad E4 gene complex, which has previously been demonstrated to support the long-term survival of ECs through sustained Akt phosphorylation [26 (link)]. The cells were maintained within a trigas humidified Heracell 150i incubator (Thermo Scientific, Waltham, MA, USA) at 37 °C and 5% CO₂/O₂. For experiments to inhibit the mTOR’s kinase activity, the cells were pre-treated with rapamycin (Cell Signaling Technology, Danvers, MA, USA) at a concentration of 100 nM for 24 h prior to stretch.

Lai M.W., Chow N., Checco A., Kunar B., Redmond D., Rafii S, & Rabbany S.Y. (2022). Systems Biology Analysis of Temporal Dynamics That Govern Endothelial Response to Cyclic Stretch. Biomolecules, 12(12), 1837.

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Culturing Human and Murine Cells for Toxoplasma Assays

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Human foreskin fibroblasts (HFF) and murine macrophages were obtained from American Type Culture Collection (ATCC). All cell lines and parasite strains were maintained in D10 media which consisted of DMEM media (Lonza) supplemented with 10% heat inactivated Hyclone bovine serum (GE Healthcare Life Sciences), HyClone 2 mM L-glutamine (GE Healthcare Life Sciences), 100 μg/mL penicillin and streptomycin (Corning), 20% Medium 199 (Corning) and gentamicin sulfate (Corning) at 37°C with 5% CO₂. Type I strain of T. gondii constitutively expressing red fluorescent dimerized Tomato (“RH-dTom”) and a type II strain, PRU expressing the same fluorophore (“PRU-dTom”) were used in assays.

Sanford A., Schulze T., Potluri L., Hemsley R., Larson J., Judge A., Zach S., Wang X., Charman S., Vennerstrom J, & Davis P. (2018). Novel Toxoplasma gondii Inhibitor Chemotypes. Parasitology international, 67(2), 107-111.

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Isolation and Stimulation of Human Aortic Endothelial Cells

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HAECs were isolated from aortic explants of heart transplant donors in association with the UCLA Transplant Program. Isolation of HAECs involves selective enzymatic digestion and results in high purity of the ECs as previously described (Navab et al., 1988 (link); Romanoski et al., 2011 (link)). Confluent HAECs in Medium 199 (Corning) supplemented with 1% heat-inactivated fetal bovine serum and 1% antibiotics were treated with Ox-PAPC as described in a previous publication (Romanoski et al., 2010 (link)), 10 ng/ml recombinant human TNF, or 10 ng/ml IL-1β. The dose of Ox-PAPC (40–50 µg/ml) used for these experiments was chosen based on experiments from our laboratory and others showing consistent atherogenic effects at this concentration (Lee et al., 2012 (link)). Although Ox-PAPC can have antiinflammatory action on bacterial products at much lower concentrations (Oskolkova et al., 2010 (link)), aortic extracts of oxidized phospholipid at the concentration used in this study have been shown to have proinflammatory effects on ECs (Subbanagounder et al., 2000 (link); Oskolkova et al., 2010 (link)). For rescue experiments, confluent HAECs were pretreated with 10 µM Stattic (Selleck Chemicals) or vehicle control for 30 min; the cells were then treated with control media or media containing 10 ng/ml TNF or 40 µg/ml Ox-PAPC in the presence or not of the inhibitor for 4 h.

Briot A., Civelek M., Seki A., Hoi K., Mack J.J., Lee S.D., Kim J., Hong C., Yu J., Fishbein G.A., Vakili L., Fogelman A.M., Fishbein M.C., Lusis A.J., Tontonoz P., Navab M., Berliner J.A, & Iruela-Arispe M.L. (2015). Endothelial NOTCH1 is suppressed by circulating lipids and antagonizes inflammation during atherosclerosis. The Journal of Experimental Medicine, 212(12), 2147-2163.

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