L 15 media

Manufactured by Thermo Fisher Scientific

Sourced in United States

L-15 media is a cell culture medium designed for the growth and maintenance of a variety of cell types. It is a balanced salt solution that provides the necessary nutrients and osmotic environment for cell survival and proliferation. The core function of L-15 media is to support the in vitro culture of cells.

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Lab products found in correlation

Close spelling variants of this product

Leibovitz’s L-15 media (38 citations) Leibovitz L-15 media (33 citations) Leibovitz’s L-15 cell culture media (3 citations) L-15 imaging media (2 citations) L-15 glutamax media (2 citations) Leibovitz L-15 and McCoy 5A (LM) media (2 citations)

65 protocols using l 15 media

Hypoxic Exposure of Ovarian Cancer Cells

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The human OvCa cell lines, OVCAR-3 (HTB-161), SW 626 (HTB-78), and TOV-112D (CRL-11731), were purchased from the American Tissue Culture Collection (ATCC, Manassas, VA). OVCAR-3 cells were grown in RPMI-1640 media supplemented with 0.01 mg/ml bovine insulin, and 20% fetal bovine serum (FBS) (Fisher Scientific, Pittsburgh, PA). SW 626 cells were grown in L-15 media (Fisher Scientific, Pittsburgh, PA) supplemented with 10% FBS. TOV-112D cells were maintained in a 1:1 mixture of MCDB 105 medium containing 1.5 g/L sodium bicarbonate and Medium 199 containing 2.2 g/L sodium bicarbonate, supplemented with 15% FBS and penicillin/streptomycin antibiotic solution (Fisher Scientific, Pittsburgh, PA). All cell lines were maintained in an incubator at 37 °C and 5% CO₂.
For hypoxic exposure, cells were incubated in a humidified atmosphere of 1% O₂, 5% CO₂ and 94% N₂ at 37 °C for 3–12 h in a special hypoxia environmental chamber (Stem Cell technology, Canada). Alternatively, for cells in a standard cell culture incubator, CoCl₂ (150 μM) (Fisher Scientific, Pittsburgh, PA) was added to the medium to mimic a hypoxic environment.

Singh S.K., Mishra M.K, & Singh R. (2019). Hypoxia-inducible factor-1α induces CX3CR1 expression and promotes the epithelial to mesenchymal transition (EMT) in ovarian cancer cells. Journal of Ovarian Research, 12, 42.

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Culturing OVCAR-3 and SW 626 Cell Lines

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The OvCa cell lines, OVCAR-3, and SW 626 were purchased from American Tissue Culture Collection (ATCC, Manassas, VA, USA). Following to the ATCC cell culture method with a small modification, OVCAR-3 cells were grown in RPMI media supplemented with 20% Fetal Bovine Serum (FBS) (Fisher scientific, PA, USA), 0.01 mg/ml bovine insulin, 1% of Non-essential amino acid solution and 1% mL of penicillin/streptomycin solution (Fisher Scientific, PA, USA). However, SW 626 cells were grown in L-15 media (Fisher Scientific, PA, USA), supplemented with 10% FBS, and penicillin/streptomycin solution. All cell cultures were maintained in a humidified incubator at 37 °C and 5% CO₂.

Ukwade C.E., Ebuehi O.A., Adisa R.A., Singh S.K, & Singh R. (2020). Anti-proliferative activities of Byrsocarpus coccineus Schum. and Thonn. (Connaraceae) using ovarian cancer cell lines. Journal of Ovarian Research, 13, 83.

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Multimodal Live-Cell Imaging Protocol

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Cell images were taken with a Leitz-Wetzlar Type 307 microscope equipped with a 12-bit CCD camera and a Nikon Eclipse Ti2 microscope. Fluorescence channels were merged using ImageJ software. Timelapse imaging was conducted with HeLa cells stably expressing GFP-tagged GalT and HeLa cells stably expressing GFP-tubulin on an Olympus FluoView1000 using a 20x/0.95 NA water objective. Z-stack slices were optimized for the objective at 1.18 mm thickness. For the duration of the imaging, cells were on a 37 °C heat plate and maintained in L-15 media (Thermo Scientific, Waltham, MA) supplemented with 10% fetal bovine serum. Cells transfected with iRFP-tagged plasmids were incubated with 5 μM biliverdin (Cayman Chemical, Ann Arbor, MI) for 30 min prior to imaging.

Walton K., Nawara T.J., Angermeier A.R., Rosengrant H., Lee E., Wynn B., Victorova E., Belov G, & Sztul E. (2023). Site-specific phosphorylations of the Arf activator GBF1 differentially regulate GBF1 function in Golgi homeostasis and secretion versus cytokinesis. Scientific Reports, 13, 13609.

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Tamoxifen-Induced Lineage Tracing

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Homozygous Venus::HES5 knock-in females were mated with R26R-H2B::mCherry Sox1Cre:ERT2 males and E0.5 was considered as midday on the day a plug was detected. Intra-peritoneal injection of pregnant females with 2.5 mg Tamoxifen (Sigma) was performed 18 h prior to embryo dissection. Whole embryos were screened for H2B::mCherry expression using Fluar ×10/0.5NA objective on a Zeiss LSM880 confocal microscope and the trunks of positive embryos were embedded in 4% low-gelling temperature agarose (Sigma) containing 5 mg/ml glucose (Sigma). 200 μm transverse slices of the trunk around the forelimb region were obtained with the Leica VT1000S vibratome and released from the agarose. Embryo and slice manipulation was performed in phenol-red-free L-15 media (Thermo Fisher Scientific) on ice and the vibratome slicing was performed in chilled 1xPBS (Thermo Fisher Scientific).

Manning C.S., Biga V., Boyd J., Kursawe J., Ymisson B., Spiller D.G., Sanderson C.M., Galla T., Rattray M, & Papalopulu N. (2019). Quantitative single-cell live imaging links HES5 dynamics with cell-state and fate in murine neurogenesis. Nature Communications, 10, 2835.

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Tumor Cell Isolation for Single-Cell RNA-Seq

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Micro-dissected lung tumors were dissociated with collagenase IV, dispase, and trypsin at 37°C for 30 minutes as previously described^{118 (link)}. The digestion buffer was then neutralized with cold L-15 media (Thermo Fisher Scientific: 21083027) containing 5% FBS (Gemini Bio) and DNaseI (Sigma-Aldrich: DN25). Dissociated cells were treated with ACK Lysis Buffer (Thermo Fisher Scientific: A1049201) and resuspended in PBS containing 2 mM EDTA (Promega: V4233), 2% FBS, and DNase I. For the isolation of neoplastic cells, dissociated cells were stained with DAPI and antibodies against CD45 (BioLegend: 103112; 1:800 dilution), CD31 (BioLegend: 102402; 1:800 dilution), F4/80 (BioLegend: 123116; 1:800 dilution), and Ter119 (BioLegend: 116212; 1:800 dilution) to exclude hematopoietic and endothelial cells. FACSAria™ sorters (BD Biosciences) were used for cell sorting. For sorting of total cells within tumors for single-cell RNA-seq, dissociated cells were stained with DAPI only to exclude dead cells. Representative gating scheme included in Supplementary Fig. 20a.

Murray C.W., Brady J.J., Han M., Cai H., Tsai M.K., Pierce S.E., Cheng R., Demeter J., Feldser D.M., Jackson P.K., Shackelford D.B, & Winslow M.M. (2022). LKB1 drives stasis and C/EBP-mediated reprogramming to an alveolar type II fate in lung cancer. Nature Communications, 13, 1090.

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Breast Cancer Cell Line Characterization

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Two breast cancer cell lines, MCF-7 and MDA-MB-231, were obtained from the American Type Tissue Culture Collection (ATCC, Manassas, VA, USA). MCF-7 cells were cultured in Dulbecco’s modified Eagle medium (DMEM; Sigma-Aldrich, St Louis, MO, USA) supplemented with 10% (v/v) foetal bovine serum (FBS) and penicillin and streptomycin (Sigma-Aldrich). MDA-MB-231 cells were maintained in L-15 media (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS and penicillin and streptomycin. p53 knockout (KO) and p53/c-Myc double-knockout (DKO) MCF-7 cells were generated by CRISPR‒Cas9 genome editing, as previously described^{57 (link)}.

Aersilan A., Hashimoto N., Yamagata K., Yokoyama M., Nakayama A., Shi X., Nagano H., Sakuma I., Nohata N., Kinoshita T., Seki N., Rahmutulla B., Kaneda A., Zhahara S.N., Gong Y., Nishimura M., Kawauchi S., Kawakami E, & Tanaka T. (2022). MicroRNA-874 targets phosphomevalonate kinase and inhibits cancer cell growth via the mevalonate pathway. Scientific Reports, 12, 18443.

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Primary Zebrafish Motor Neuron Culture

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Primary zebrafish motor neurons were cultured from pooled 18hpf male and female Tg(mnx1:mCherry-PA-Rac1);mitfa^w2/w2;mpv17^a9/a9 and Tg(mnx1:mCherry-PI-Rac1);mitfa^w2/w2;mpv17^a9/a9 fish at room temperature in Leibovitz’s L-15 media (Thermo Fischer 21083027) supplemented with 2% fetal bovine serum and 100U/mL penicillin/streptomycin.

Harris J.M., Wang A.Y., Boulanger-Weill J., Santoriello C., Foianini S., Lichtman J.W., Zon L.I, & Arlotta P. (2020). Long-range optogenetic control of axon guidance overcomes developmental boundaries and defects. Developmental cell, 53(5), 577-588.e7.

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Cell Line Culture and Characterization

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MCF‐7, MDA‐MB‐231 and SK‐Br‐3 cells were from the cell bank of the Chinese Academy of Sciences (Shanghai, China). MCF‐7 and SK‐Br‐3 cells were cultured in MEM media (Thermo Fisher, Waltham, MA, USA), and MDA‐MB‐231 cells were cultured in L15 media (Thermo Fisher), supplemented with 10% FBS (Thermo Fisher), 100 units/mL penicillin and 100 μg/mL streptomycin at 37°C in a humidified incubator with 5% CO₂. Before the study, the cells were passaged for 6 generations. The identity of the cell lines was determined by short tandem repeat profiling.

Sun W., Wang L., Luo J., Zhu H, & Cai Z. (2018). Ambra1 modulates the sensitivity of breast cancer cells to epirubicin by regulating autophagy via ATG12. Cancer Science, 109(10), 3129-3138.

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Neutrophil Chemotaxis Assay using Microfluidics

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Priming of each device commenced by pipetting 20 μL of an fMLF-based chemoattractant solution through the loading port. For this, fMLF (Sigma Aldrich, stored in stock solutions of 1 μM in DMSO) was diluted in L-15 media (Thermo Fisher Scientific) containing 20% hiFBS to a final concentration of 100 nM. The device was then placed in a vacuum desiccator connected to house vacuum (27 inHg) for 10 min. Upon removal the device was rested for an additional 10 min, until the chemoattractant filled entirely the straight migration channels, connected orthogonally to the central loading channel. The device was then washed twice, by pipetting 200 μL of 10 μg/mL fibronectin (Sigma Aldrich) diluted in L-15 media containing 20% hiFBS into the loading port. These washing steps removed the chemoattractant from the central loading channel and its passive diffusion from the migration channels into the central loading channel established a chemoattractant gradient. To prevent evaporation, 3 mL of L-15 media were added to the glass-bottomed dish to cover the device.

Hadjitheodorou A., Bell G.R., Ellett F., Irimia D., Tibshirani R., Collins S.R, & Theriot J.A. (2023). Leading edge competition promotes context-dependent responses to receptor inputs to resolve directional dilemmas in neutrophil migration. Cell systems, 14(3), 196-209.e6.

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Tumor Growth in NOD SCID Mice

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Cells were injected into the hind flank of NOD.CB17-Prkdc^scid/J mice(Taconic, Germantown, NY) in a solution containing 50% L-15 media (ThermoFisher) with 1mM HEPES (MilliporeSigma) and 50% matrigel (BD). 10 tumors were used for each group in each experiment. All studies were approved by the University of South Florida IACUC (#IS00004987). PLX 4720 was given using formulated chow (Research Diets, New Brunswick, NJ) while panobinostat and PCI-34051 were given by i.p. injections for the duration of the experiment. Weight and tumor size were measured with calipers and were monitored 3 times weekly.

Emmons M.F., Faião-Flores F., Sharma R., Thapa R., Messina J.L., Becker J.C., Schadendorf D., Seto E., Sondak V.K., Koomen J.M., Chen Y.A., Lau E.K., Wan L., Licht J.D, & Smalley K.S. (2019). HDAC8 regulates a stress response pathway in melanoma to mediate escape from BRAF inhibitor therapy. Cancer research, 79(11), 2947-2961.

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