Mccoy s 5a

Manufactured by American Type Culture Collection

Sourced in United States

McCoy's 5A is a cell culture medium designed for the growth and maintenance of various cell lines. It provides the necessary nutrients and growth factors to support the culture of cells in vitro. The medium composition is standardized to ensure consistent performance and reliable results.

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Lab products found in correlation

Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (10 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (9 mentions) Dulbecco modified eagle medium (dmem), by American Type Culture Collection (4 mentions) Hct116 cell, by American Type Culture Collection (4 mentions) Fetal bovine serum (fbs), by Corning (3 mentions) Ht 29, by American Type Culture Collection (3 mentions) Hct116, by American Type Culture Collection (3 mentions) Fetal bovine serum (fbs), by American Type Culture Collection (3 mentions) Mccoy s 5a, by Thermo Fisher Scientific (3 mentions) Rpmi 1640, by American Type Culture Collection (3 mentions) Rpmi 1640, by Corning (2 mentions) Mycoalert mycoplasma detection kit, by Lonza (2 mentions) Hek293t, by American Type Culture Collection (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Sodium pyruvate, by Thermo Fisher Scientific (2 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by Lonza (2 mentions) Caco 2, by American Type Culture Collection (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Recombinant human ifn γ, by Merck Group (2 mentions)

Spelling variants of this product

McCoy’s 5A medium (115 citations) McCoy's 5a Modified Medium (32 citations) McCoy’s 5A media (9 citations) McCoy’s 5A culture medium (2 citations) McCoy’s 5A growth medium (2 citations) McCoy’s 5a Medium Modified (ATCC® 30-2007™) (2 citations)

33 protocols using mccoy s 5a

Inhibition of HER2-Expressing Cell Lines

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Example 11

The antibodies from the D-series of immunizations were tested for their ability to inhibit growth of the high-HER2 expressing cell lines SKBR3 and BT474. SKBR3 and BT474 cells were plated in ½-area 96-well plates at a density of 120 k/well and 240 k/well, respectively. SKBR3 cells were cultured in 100 μL McCoy's 5A (ATCC) 10% FBS, 1% penicillin-streptomycin and 2 mM L-glutamine. BT474 cells were maintained in 100 μL at 37° C. in 5% CO₂in 50:50 DMEM: F12, 10% FBS, 1% penicillin-streptomycin and 2 mM L-glutamine. After cells were allowed to attach for 4 hours, purified antibodies listed in FIG. 13 were added to final concentrations of 1.0, 3 and 10 μg/mL. 4D5 included as a positive control. The cells were allowed to grow for 3 days before cell growth was assessed by the XTT assay (Sigma) per the manufacturer's instructions. The difference in absorbance at 492 nm and 690 nm was taken as proportional to the number of cells in each well. These results (FIG. 13) suggest that D3.4 and to some degree D4.1 inhibit the growth of SKBR3 cells but not BT474. This difference in reactivity towards two cells lines with near equally high levels of HER2 expression may be explained by the fact that SKBR3 is know to shed greater levels of HER2 extracellular domain into the media and therefore may be more dependent on p95 retained in the cell.

US10273308B2. Methods of producing antibodies specific for p95 (2019-04-30). Laboratory Corporation of America Holdings [US]. Inventors: Jeff Sperinde [US], John William Winslow [US], Xueguang Jin [US], Gerald J. Wallweber [US].

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Cell Culture Protocols for U2OS and HEK293T

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U2OS cells were grown in McCoy's 5A (ATCC) supplemented with 10% fetal bovine serum (Gibco). HEK293T cells were grown in DMEM (Gibco) supplemented with 10% fetal bovine serum (Gibco).

Luo K., Deng M., Li Y., Wu C., Xu Z., Yuan J, & Lou Z. (2015). CDK-mediated RNF4 phosphorylation regulates homologous recombination in S-phase. Nucleic Acids Research, 43(11), 5465-5475.

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Culturing HT29 and LLC Cell Lines

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HT29 cells (ATCC) were cultured in McCoy’s 5A (ATCC) medium, supplemented with 10% FBS (12 (link)). Highly angiogenic and VEGFA producing type LLC cells (clone D122-96) was provided by Dr. L. Eisenbach (Weizman Institute of Science, Rehovot, Israel) and were cultured in DMEM (ATCC) containing 10% FBS (15 (link)).

Sarkar C., Chakroborty D., Dasgupta P.S, & Basu S. (2015). Dopamine is a safe anti-angiogenic drug which can also prevent 5-fluorouracil induced neutropenia. International journal of cancer. Journal international du cancer, 137(3), 744-749.

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Cultivation of OVCAR3 and HT-29 Cell Lines

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Both OVCAR3 and HT‐29 were obtained from ATCC. Cell lines were cultured at 37°C with 5% CO₂ in ATCC‐recommended media with 10%‐20% heat inactivated fetal bovine serum (FBS; Corning) supplemented with 2 mmol/L L‐glutamine (L‐glut; MP Biomedicals LLC). OVCAR3 was grown in RPMI 1640 (Corning), 20% FBS, and L‐glut. HT‐29 was grown in McCoy's 5A (ATCC), 10% FBS, and L‐glut. Cells were maintained at optimal confluency. All cell lines, periodically tested, were negative for mycoplasma contamination, using the MycoAlert Mycoplasma Detection Kit from Lonza. The OVCAR3 cell line was sent in and cell line validated by ATCC with the Short Tanden Repeat (STR) profiling test (Manassas, VA).

Minnix M., Li L., Yazaki P., Chea J., Poku E., Colcher D, & Shively J.E. (2020). Improved targeting of an anti‐TAG‐72 antibody drug conjugate for the treatment of ovarian cancer. Cancer Medicine, 9(13), 4756-4767.

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Cell Culture Protocols for KBM7, HEK293T, and HCT116

Cited 1 time

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KBM7 cells were obtained from Thijn Brummelkamp (Carette et al., 2009 (link), 2011 (link)) and were cultured at 37°C in 5% CO₂ in Iscove's Modified Dulbecco Medium (IMDM) (Gibco, Thermo Fisher Scientific, Waltham, MA) supplemented with 10% heat inactivated fetal bovine serum (Gibco), and penicillin/streptomycin at final concentrations of 100 U/mL and 100 μg/mL, respectively (P/S) (Corning Inc., Corning, NY). HEK293T and HCT116 were obtained from ATCC (Manassas, VA). HEK293T were cultured at 37°C in 5% CO₂ in Dulbecco’s Modified Eagle Medium (DMEM) (ATCC) supplemented with 10% fetal bovine serum (FBS) (ATCC), and P/S. HCT116 were cultured at 37°C in 5% CO₂ in McCoy's 5A (ATCC) supplemented with 10% FBS, and P/S. Cell lines were used at low passage numbers from primary stocks and were not further authenticated or tested for mycoplasma.

Chidley C., Trauger S.A., Birsoy K, & O'Shea E.K. (2016). The anticancer natural product ophiobolin A induces cytotoxicity by covalent modification of phosphatidylethanolamine. eLife, 5, e14601.

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Characterization of PIK3CA mutant cell lines

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All cell lines were purchased from ATCC, DSMZ, JCRB or MD Anderson (Minna Lab), except for TM00244, which was purchased as a PDX from Jackson Labs. We generated a stable cell line from TM00244 and confirmed that it retained the PIK3CA mutation. All cell lines were maintained at 37°C in 5% CO₂ in RPMI (Gibco; Catalog No. 11875–093), DMEM (Gibco; Catalog No. 11995–065), EMEM (ATCC; Catalog No. 30–2003), or McCoy’s 5A (ATCC; Catalog No. 30–2007) media and supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, and 1% glutamine, except for RERFLCSQ1 and LOU-NH91, which were supplemented with 10% or 20% heat-inactivated fetal bovine serum, respectively. Cells were used within 20 passages from thaw for experiments and were STR profiled and routinely tested for mycoplasma (Lonza MycoAlert; Catalog No. LT07–703) every 6 months. For in vitro use, CC-115 (Selleck Chemicals; Catalog No. S7891) was dissolved in DMSO and clinical grade carboplatin (Rutgers Cancer Institute of New Jersey Pharmacy) was diluted in media. The molecular structure of CC-115 was previously published (31 (link)).

Castellano G.M., Zeeshan S., Garbuzenko O.B., Sabaawy H.E., Malhotra J., Minko T, & Pine S.R. (2022). Inhibition of mTORC1/2 and DNA-PK via CC-115 Synergizes with Carboplatin and Paclitaxel in Lung Squamous Cell Carcinoma. Molecular cancer therapeutics, 21(9), 1381-1392.

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Comparison of Three Osteosarcoma Cell Lines

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Three human osteosarcoma cell lines were used: U2OS (ATCC HTB-96), and SaOS2 (ATCC HTB-85), and LM7, which was a gift from Dr. Eugenie Kleinerman (Children’s Cancer Hospital, the University of Texas MD Anderson Cancer Center, Houston, TX). The LM7 cell line was created by passaging 10⁶ SaOS2 cells through lungs of athymic mice 7 times^{19 (link),20 (link)}. U2OS was grown on McCoy’s5A (ATCC cat. 30–2007) with 10% FBS and 1% penicillin-streptomycin (Gibco cat. 10378–016), at 37°C in 5% CO2. SaOS2 was grown in similar media and growth conditions except with 15% FBS. LM7 was grown on DMEM (Lonza cat. 12–614F), with 10% FBS, 1mM non-essential amino acids (Gibco cat. 11140–050), 1mM sodium pyruvate (Gibco cat. 11360–070), 2mM penicillin-streptomycin with glutamine (Gibco cat. 10378–016), and 2X MEM vitamin solution (Gibco cat. 11120–052).

Loh A.H., Stewart E., Bradley C.L., Chen X., Daryani V., Stewart C.F., Calabrese C., Funk A., Miller G., Karlstrom A., Krafcik F., Goshorn D.R., Vogel P., Bahrami A., Shelat A, & Dyer M.A. (2018). Combinatorial Screening Using Orthotopic Patient Derived Xenograft-Expanded Early Phase Cultures of Osteosarcoma Identify Novel Therapeutic Drug Combinations. Cancer letters, 442, 262-270.

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Cell Culture Protocol for Cancer Research

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All cell lines and primary cells were cultured at 37 °C, 5% CO2. HEK-293T, SK-MEL-5 and CHL-1 were cultured on DMEM 10% FBS, 1% Pen-Strep; HT-1376, Caco-2, SK-MEL-2 and LS174T were cultured in EMEM 20% FBS, 1% Pen-Strep; BxPC-3, MM415, MM485, AsPC-1, DLD-1 KRAS^WT/−, and DLD-1 KRAS^G13D/− were cultured in RMPI-1460 (ATCC Modification) 10% FBS, 1% Pen-Strep; T24 and Capan-2 were cultured in McCoy’s 5A (Modified) 10% FBS, 1% Pen-Strep. CHL-1, HT-1376, Caco-2, Capan-2, SK-MEL-2, SK-MEL-5, LS174T, AsPC1 and T24 lines were purchased authenticated from ATCC. DLD-1 KRAS^WT/−, and DLD-1 KRAS^G13D/− were purchased from Horizon Discovery, Ltd. MM415 and MM485 were authenticated by polymorphic short tandem repeat (STR) loci profiling by Promega. HEK-293T, CHL-1, SK-MEL-5, BxPC-3, AsPC-1, LS174T, HT-1376 and T24 are derived from female individuals. Caco-2, DLD-1, MM415, MM485, Capan-2 and SK-MEL-2 are derived from male individuals.
Primary human melanocytes were isolated from fresh biopsy samples approved by the Stanford University IRB and were propagated up to eight passages in Media 254 with HMGS supplement (Thermo Fisher Scientific Scientific) and 1% Pen-Strep. All primary human melanocytes were male. All cell lines and primary cells were confirmed to be mycoplasma free bimonthly using the MycoAlert mycoplasma detection kit (Lonza).

Kovalski J.R., Bhaduri A., Zehnder A.M., Neela P.H., Che Y., Wozniak G.G, & Khavari P.A. (2019). The Functional Proximal Proteome of Oncogenic Ras Includes mTORC2. Molecular cell, 73(4), 830-844.e12.

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Vero and 13762 MAT B III Cell Culture

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Vero and 13762 MAT B III cells were purchased from the American Type Culture Collection (Manassas, VA). 13762 MAT B III cells were maintained in McCoy’s 5A (ATCC) supplemented with 10% fetal bovine serum (FBS) (HyClone, Logan, UT). Vero cells were maintained in Dulbecco’s Modiﬁed Eagle’s medium (DMEM) (HyClone) supplemented with 10% FBS (HyClone).
Maraba and MG1 were used as previously described.^{2 (link)} MRB LCMV GP was constructed as previously described with VSV.^{10 (link)} VSVΔ51 and VSV LCMV GP were used as previously described.^{1 (link),10 (link)}

Evgin L., Ilkow C.S., Bourgeois-Daigneault M.C., de Souza C.T., Stubbert L., Huh M.S., Jennings V.A., Marguerie M., Acuna S.A., Keller B.A., Lefebvre C., Falls T., Le Boeuf F., Auer R.A., Lambris J.D., McCart J.A., Stojdl D.F, & Bell J.C. (2016). Complement inhibition enables tumor delivery of LCMV glycoprotein pseudotyped viruses in the presence of antiviral antibodies. Molecular Therapy Oncolytics, 3, 16027-.

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Cell Culture Procedures for Cancer and Primary Cell Lines

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HCT 116, MDA-MB-231 and HCC1806 cells were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). EW8, Rh41 and HCC1806 cells were maintained in RPMI 1640 (Lonza, Portsmouth, NH, USA) supplemented with 10% fetal bovine serum (FBS; Life Technologies), TC-71 was maintained in IMDM (Lonza) supplemented with 10% FBS and ITS (Life Technologies), HCT 116 cells were maintained in McCoy's 5A (ATCC) supplemented with 10% FBS, and MDA-MB-231 cells were maintained in DMEM (Lonza) supplemented with 10% FBS at 37 °C with 5% CO₂.
Primary MEF cells were derived from E13.5 or E14.5 embryos. The p53^−/− and ARF^−/− MEF cells are generous gifts from Dr. Martine F. Roussel (St. Jude Children's Research Hospital). MEF cells were maintained in DMEM (Lonza) that contained 10% FBS, 1 × MEM non-essential amino acid, 55 μM 2-mercaptoethanol and 10 μg/ml gentamicin (Life Technologies) at 37 °C with 5% CO₂. Early passage (P2-P5) MEF cells were used for the experiments. Exceptions were p53^−/− and ARF^−/− MEF cells.

Ogi H., Sakuraba Y., Kitagawa R., Xiao L., Shen C., Cynthia M.A., Ohta S., Arnold M.A., Ramirez N., Houghton P.J, & Kitagawa K. (2015). The oncogenic role of the cochaperone Sgt1. Oncogenesis, 4(5), e149-.

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