A2058

Manufactured by American Type Culture Collection

Sourced in United States, Germany, China

The A2058 is a laboratory equipment used for the cultivation and maintenance of cell cultures. It provides a controlled environment for the growth and propagation of various cell lines. The core function of the A2058 is to support the culturing and preservation of cells in a consistent and reliable manner.

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Lab products found in correlation

139 protocols using a2058

Culturing Melanoma Cell Lines and Melanocytes

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Human melanoma cell line A375 (malignant, CRL-1619), G361 (malignant, CRL-1424), A2058 (metastatic, CRL-11147), SK-MEL-3 (metastatic, HTB-69), normal human primary epidermal melanocytes (PCS-200-013) were all purchased from American Type Culture Collection (ATCC). A375, A2058 cell lines were cultured in DMEM (ATCC) supplemented with 10% fetal bovine serum (FBS, Gibco). G361, SK-MEL-3 cell lines were cultured in McCoy's 5A (ATCC) supplemented with 10% and 15% fetal bovine serum (FBS, Gibco), respectively. Normal human primary epidermal melanocytes were grown in dermal cell basal media (ATCC, PCS-200-030) supplemented with adult melanocyte growth kit (ATCC, PCS-200-042) components. All cells were grown in a humidified incubator at 37°C, 5% CO₂ atmosphere. After 1-2 days, cells were chosen to perform calcium imaging or other functional experiments.

Zheng J., Liu F., Du S., Li M., Wu T., Tan X, & Cheng W. (2019). Mechanism for Regulation of Melanoma Cell Death via Activation of Thermo-TRPV4 and TRPV2. Journal of Oncology, 2019, 7362875.

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Cell Culture Conditions for Cancer and Normal Cell Lines

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Human hepatocellular liver carcinoma (HepG2; ATCC^® HB-8065™) were cultured in EMEM medium, human melanoma cells (A2058; ATCC^®CRL-11147^TM) were cultured in DMEM, human keratynocytes (HaCaT, CLS n. 300493) were cultured in DMEM medium, adenocarcinomic human alveolar basal epithelial cells (A549; ATCC^®CL-185^TM) were cultured in F-12K medium and human normal lung fibroblasts (MRC-5; ATCC^® CCL-171™) were cultured in EMEM medium. The media were supplemented with 10% fetal bovine serum, 50 U·mL⁻¹ penicillin and 50 μg·mL⁻¹ streptomycin. Human cell lines A2058, A549, MRC-5 and HepG2 were bought at ATCC (https://www.lgcstandards-atcc.org/ accessed on 1 June 2019). The HACAT cells were bought at the CEINGE facility cell bank (https://www.ceinge.unina.it/en/cell-cultures accessed on 1 June 2019).

Riccio G., Martinez K.A., Martín J., Reyes F., D’Ambra I, & Lauritano C. (2022). Jellyfish as an Alternative Source of Bioactive Antiproliferative Compounds. Marine Drugs, 20(6), 350.

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Cell Line Validation and Culture

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Human melanoma cell lines A2058 and SK-MEL-28, and Ewing’s sarcoma cell line A673 were obtained from ATCC. HEK293T, CCC-HEL-1, HL-7702, and Mel JuSo were tested for eight STR loci and amegloenin gene. All cell lines were tested negative for mycoplasma. HL-7702 cells were cultured in RPMI medium and the other cells were cultured in DMEM medium supplemented with 10% FBS at 37°C with 5% CO₂ in a humidified atmosphere for less than 6 months after resuscitation.

Jalal S., Zhang T., Deng J., Wang J., Xu T., Zhang T., Zhai C., Yuan R., Teng H, & Huang L. (2022). β-elemene Isopropanolamine Derivative LXX-8250 Induces Apoptosis Through Impairing Autophagic Flux via PFKFB4 Repression in Melanoma Cells. Frontiers in Pharmacology, 13, 900973.

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Cell Line Cultivation and Validation

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The ESFT cell line A673, the heptacellular carcinoma cell line HepG2, the normal liver cell line THLE2 and the melanoma cell line A2058 were purchased from ATCC, and MHH-ES-1 was purchased from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ, Germany). These cells were tested by the cell banks for eight STR loci and the amelogenin gene. A673, MHH-ES-1 and A2058 were grown in DMEM supplemented with 10% fetal bovine serum (FBS). HepG2 was grown in MEM supplemented with 10% FBS. THLE2 was grown in Bronchial Epithelial Cell Growth Medium (BEGM) supplemented with 5 ng/mL EGF, 70 ng/mL phosphoethanolamine and 10% FBS. The flasks used for THLE2 were precoated with a mixture of 0.01 mg/mL fibronectin, 0.03 mg/mL bovine collagen type I and 0.01 mg/mL bovine serum albumin dissolved in BEBM medium. All the cell lines were incubated at 37°C with 5% CO₂. Cell lines purchased were passaged less than 30 passages after resuscitation.

Wu D., Lv D., Zhang T., Guo L., Ma F., Zhang C., Lv G, & Huang L. (2018). Antitumor effects of β-elemene via targeting the phosphorylation of insulin receptor. Endocrine-Related Cancer, 26(2), 187-199.

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Cell Line Repository for Cancer Research

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Human lung cancer cell line A549 (ATCC no. CCL-185), human melanoma cell lines HS294T (ATCC no. HTB-140), A2058 (ATCC no. CRL-11147), SK-MEL-2 (ATCC no. HTB-68), human monocyte cell line THP-1 (ATCC no. TIB-202), monkey kidney cell line CV-1 (ATCC no. CCL-70), and mouse melanoma cell line B16-F10 (ATCC no. CRL-6475) were obtained from the American Type Culture Collection (ATCC, Manassas, VA). A highly metastatic human prostate cancer cell line PC-3MM2 was generously obtained from Isaiah J. Fidler, MD Anderson Cancer Center (Houston, TX). The human bladder cancer EJ cells were provided and authenticated by A.G. Eliopoulos, University of Crete Medical School and Laboratory of Cancer Biology (Heraklion, Crete, Greece). B16-OVA, a mouse melanoma cell line expressing chicken ovalbumin, was kindly provided by Professor Richard Vile at Mayo Clinic (Rochester, MN). Jurkat cells, immortalized human T-lymphocytes, were kindly provided by Dr. Pentti Tienari, Program of Molecular Neurology, University of Helsinki (Helsinki, Finland). All cell lines were cultured under recommended conditions.

Hirvinen M., Capasso C., Guse K., Garofalo M., Vitale A., Ahonen M., Kuryk L., Vähä-Koskela M., Hemminki A., Fortino V., Greco D, & Cerullo V. (2016). Expression of DAI by an oncolytic vaccinia virus boosts the immunogenicity of the virus and enhances antitumor immunity. Molecular Therapy Oncolytics, 3, 16002-.

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Melanoma Cell Culture and Tissue Acquisition

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Normal human melanocytes, neonatal, lightly pigmented donor (HEMn-LP), and the malignant human melanoma cells, A375 and A2058, were purchased from ATCC (Manassas, VA, USA). The human melanoma cell line, UACC903, was a gift from Dr. Yongliang Zhao (Beijing Institute of Genomics, CAS). All melanoma cell lines were maintained in Dulbecco's modified Eagle's medium (DMEM, Gibco, Grand Island, NY, USA) supplemented with 10% (v/v) fetal bovine serum (FBS, AusGeneX, Molendinar, Qld, Australia) and penicillin (100 U/mL)-streptomycin (0.1 mg/mL) (Invitrogen, Carlsbad, CA, USA). All cells were cultured in a 37°C humidified, 5% CO₂ atmosphere incubator (Thermo Fisher Scientific, Waltham, MA, USA).
The acquisition of tissue samples was approved by the Chinese PLA General Hospital. We used primary melanoma samples, metastatic melanoma samples, and normal pigmented nevus samples to test the endogenous expression of IGFBP5. Informed consent was given by all patients examined. All human samples were collected in accordance with the “Declaration of Helsinki” (as revised in Edinburgh 2000).

Wang J., Ding N., Li Y., Cheng H., Wang D., Yang Q., Deng Y., Yang Y., Li Y., Ruan X., Xie F., Zhao H, & Fang X. (2015). Insulin-like growth factor binding protein 5 (IGFBP5) functions as a tumor suppressor in human melanoma cells. Oncotarget, 6(24), 20636-20649.

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Melanoma Cell Line Authentication and Culture

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The WM852 melanoma cell line was procured from the Coriell Institute (Camden NJ, USA) and maintained in RPMI-1640 medium. The HTB63, A375 and A2058 melanoma cell lines were purchased from ATCC (Old Town Manassas, VA, USA). HTB63 melanoma cells were cultured in McCoy’s 5A medium, and both A375 and A2058 melanoma cell lines were cultured in DMEM. The culture medium for all the cell lines was supplemented with 10% FBS, antibiotics and L-glutamine. The supplier confirmed the genetic authentications of all the cell lines, and no cell line was used for more than four years. All cell lines were routinely screened for the absence of mycoplasma infection. The PKC-inhibitor (Gö6983) and ROCK-inhibitor (Y-27632) were procured from Selleckchem (Munich, Germany). The predesigned siRNAs used in this study were obtained from Thermo Fischer Scientific (Waltham, MA, USA).

Mohapatra P., Yadav V., Toftdahl M, & Andersson T. (2020). WNT5A-Induced Activation of the Protein Kinase C Substrate MARCKS Is Required for Melanoma Cell Invasion. Cancers, 12(2), 346.

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Melanocyte and Melanoma Cell Culture Protocol

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Adult human melanocytes (NHEM), derived from a healthy human subject, were purchased from Lonza (Walkersville, MD). Human melanoma cell lines (A2058, SK-MEL28, SK-MEL2, C8161 and 1205Lu) were purchased from ATCC (Manassas, VA) in a period between the year of 2014 to 2017 without further authentication. As described previously (34 (link)), frozen cells were newly thawed from low (3 (link)–10 (link)) passages. Mycoplasma was tested using PlasmoTest Mycoplasma detection kits (Invitrogen, Carlsbad, CA) and only negative cells were included in the experiments. Melanoma cells were maintained under a 5% CO₂ atmosphere in RPMI medium (Life technologies, Carlsbad, CA) supplemented with 10% heat-inactivated fetal bovine serum (FBS), 100 Units/ml penicillin and 100 μg/ml of streptomycin. Melanocytes were cultured in 254 medium supplemented with Human melanocyte growth supplements (Thermo Scientific, Waltham, MA). Cells were seeded for 24 hours, and subsequently treated with sodium ascorbate (Sigma-Aldrich, St. Louis, MO). Media were changed daily to avoid the accumulation of unabsorbed ascorbate. All cells tested negative for mycoplasma by PCR.

Mustafi S., Camarena V., Volmar C.H., Huff T.C., Sant D.W., Brothers S.P., Liu Z.J., Wahlestedt C, & Wang G. (2017). Vitamin C sensitizes melanoma to BET inhibitors. Cancer research, 78(2), 572-583.

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Melanoma Cell Culture Protocols

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Human melanoma cell lines A375 (ATCC CRL-1619), A2058 (ATCC CRL-11147), SK-MEL-2 (ATCC HTB-68), MeWo (ATCC HTB-65), murine melanoma cell lines B16 (ATCC CRL-6322), B16-F10 (ATCC CRL-6475), and primary epidermal melanocytes (ATCC PCS-200-012) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, United States). A375, A2058 and B16-F10 cells were cultured in DMEM, MeWo, SK-MEL-2, and melanocytes were cultured in MEM, and B16 was cultured in RPMI 1640, supplemented with 10% heat-inactivated fetal bovine serum, 100 U/mL penicillin and 100 μg/mL streptomycin at 37°C in a humidified 5% CO₂ incubator.

Cvetanova B., Shen Y.C, & Shyur L.F. (2019). Cumingianoside A, a Phyto-Triterpenoid Saponin Inhibits Acquired BRAF Inhibitor Resistant Melanoma Growth via Programmed Cell Death. Frontiers in Pharmacology, 10, 30.

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Transient Transfection of Cell Lines

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HEK293 cells were grown in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS). Transient transfections were performed with Transfectin reagent (Bio-Rad, 1703352). The lung cancer cell lines A549 (ATCC CCL-185) and H1666 (ATCC CRL-5885) were grown in Roswell Park Memorial Institute media (RPMI), the melanoma cell line A2058 (ATCC CRC-11147) in DMEM. The media was supplemented with 10% FBS and 10 mM N-2-hydroxyethylpiperazine-N-ethanesulfonic acid (Hepes). Quail embryo fibroblasts (QEF) were grown in Avian cell culture medium. DNA transfection was mediated using the calcium phosphate method. Primary antibodies used were the mouse anti-Rluc antibody directed against Rluc-F[1] (Chemicon, #MAB4410), mouse anti–HA-tag (Covance, MMS-10P), mouse anti-BRAF (Santa Cruz, F-7: sc-5284), mouse anti-MEK1/2 (Cell Signaling, 4684S), rabbit phospho-MEK1/2 (Ser217/221) (Cell Signaling, 9154), and rabbit anti–P-ERK1/2 (Cell Signaling, 9101).

Mayrhofer J.E., Enzler F., Feichtner A., Röck R., Fleischmann J., Raffeiner A., Tschaikner P., Ogris E., Huber R.G., Hartl M., Schneider R., Troppmair J., Torres-Quesada O, & Stefan E. (2020). Mutation-oriented profiling of autoinhibitory kinase conformations predicts RAF inhibitor efficacies. Proceedings of the National Academy of Sciences of the United States of America, 117(49), 31105-31113.

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