Crl 2336

Manufactured by American Type Culture Collection

Sourced in United States

The CRL-2336 is a cell culture flask manufactured by the American Type Culture Collection (ATCC). It is designed for the propagation and maintenance of adherent cell lines in laboratory settings. The core function of the CRL-2336 is to provide a controlled and sterile environment for the cultivation of cells.

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6 protocols using crl 2336

Cell Culture of TNBC and K562 Lineages

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TNBC cell lines HCC-1806 (ATCC, CRL 2335™), HCC-1937 (ATCC, CRL-2336™), HCC-70 (ATCC, CRL-2315™), and MDA-MB-468 (ATCC, HTB-132™), and the chronic myelogenous leukemia-derived K562 cell line (ATCC, CCL-243™) were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA). The cell lines were authenticated by DNA profiling using short tandem repeat (STR) analysis on an AmpFlSTR^® Identifier™ PCR Amplification System at the National Institute of Genomic Medicine (INMEGEN), Mexico City, Mexico. The cells were used between passage 3 and passage 15. All the cells were maintained in Roswell Park Memorial Institute (RPMI)-1640 medium (Biowest SAS, Nuaillé, France) that was supplemented with 10% heat-inactivated Fetal Bovine Serum (FBS) (Biowest SAS, Nuaillé, France), 100 U/mL penicillin, and 100 mg/mL streptomycin (Invitrogen, Carlsbad, CA, USA). The cells were maintained in a 5% CO₂ humidified atmosphere at 37 °C.

López-Mejía J.A., Tallabs-Utrilla L.F., Salazar-Sojo P., Mantilla-Ollarves J.C., Sánchez-Carballido M.A, & Rocha-Zavaleta L. (2022). c-Kit Induces Migration of Triple-Negative Breast Cancer Cells and Is a Promising Target for Tyrosine Kinase Inhibitor Treatment. International Journal of Molecular Sciences, 23(15), 8702.

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Cytotoxicity Profiling of Novel Compounds

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Human endometrial carcinoma (HeLa, ATCC, CCL-2), human lung adenocarcinoma (A549, ATCC, CCL-185), human prostate carcinoma (Du145, ATCC, HTB-81), human ductal carcinoma (HCC-1937, ATCC, CRL-2336), human breast adenocarcinoma (MCF-7, ATCC, HTB-22), and human lung fibroblasts (MRC-5, ATCC, CCL-171) were obtained from the American Type Culture Collection and maintained as exponentially growing monolayers by culturing according to the supplier’s instructions. For the cytotoxicity analysis, each cell line was exposed to the compounds (DC-AST VII and DAC-AST VII) at a final concentration of 2, 4, 8, 16, or 32 μM for 48 h. After a 48 h incubation, the cell viability was analyzed using the MTT assay (Sigma Aldrich, St. Louis, MI, ABD) according to the manufacturer’s instructions. DMSO was used as a negative control. Absorbance was measured with a microplate reader at 570 nm (Varioscan, Thermo Fisher Scientific, Waltham, MA, US). IC50 values, defined as the compound concentration at which the response level was reduced to half its maximum, were calculated using GraphPad Prism 6 (San Diego, CA, USA).

Yakubogullari N., Cagir A., Bedir E, & Sag D. (2023). Astragalus Saponins, Astragaloside VII and Newly Synthesized Derivatives, Induce Dendritic Cell Maturation and T Cell Activation. Vaccines, 11(3), 495.

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Cultivation of Breast Cancer Cell Lines

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MCF7 (HTB-22 from ATCC, Manassas, VA, USA), a model of human sporadic breast cancer cell line, luminal A, were grown in Dulbecco’s modified Eagle’s medium (DMEM) (Sigma Aldrich, Saint Louis, MO, USA, implemented with 100 mg/mL streptomycin and 100 U/mL penicillin (Sigma Aldrich) and 10% (w/v) fetal bovine serum (FBS) (Sigma Aldrich). MCF10 are a model of immortalized mammary epithelial cells (CRL-10317 from ATCC, Manassas, VA, USA). Cells were grown in MEGM, Mammary Epithelial Cell Growth Medium (Lonza, Walkersville, MD, USA), implemented with 20 ng/mL epidermal growth factor (Lonza),10 μg/mL insulin (Lonza), 0.5 μg/mL hydrocortisone (Lonza), and 100 ng/mL cholera toxin (Sigma Aldrich). The HCC1937 (CRL-2336 from ATCC, Manassas, VA, USA) are a model for hereditary breast cancer cell line that has a homozygous mutation for the BRCA1 5382C terminal; The HCC193 are a model of triple negative breast cancer. The culture medium for HCC1937 was Roswell Park Memorial Institute (RPMI) (ATCC, Manassas, VA, USA) implemented with 20% (w/v) fetal bovine serum (FBS) (Sigma Aldrich), 100 mg/mL streptomycin, and 100 U/mL penicillin (Sigma Aldrich).

Scumaci D., Olivo E., Fiumara C.V., La Chimia M., De Angelis M.T., Mauro S., Costa G., Ambrosio F.A., Alcaro S., Agosti V., Costanzo F.S, & Cuda G. (2020). DJ-1 Proteoforms in Breast Cancer Cells: The Escape of Metabolic Epigenetic Misregulation. Cells, 9(9), 1968.

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Breast Cancer Cell Line Cultivation

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The TNBC cell lines HCC1806, HCC1937, MDA-MB-157 (MDA-157), MDA-MB-231 (MDA-231), and MDA-MB-468 (MDA-468) were purchased from the American Type Culture Collection (Manassas, VA; CRL-2335, CRL-2336, HTB-24, HTB-26, and HTB-132). MX-1 was purchased from the NCI repository. All cell lines were purchased within the previous 24 months and passaged < 15 times for all experiments [Table 1]. Cells were tested biweekly for mycoplasma contamination (MycoAlert, Lonza, Basel, Switzerland). MX-1, MDA-157, MDA-231, and MDA-468 were grown in Dulbecco Modified Eagle Medium (DMEM High Glucose with GlutaMAX, Life Technologies, Carlsbad, CA) and supplemented with 1% sodium pyruvate (Life Technologies) and 10% fetal bovine serum (FBS) (Premium Select, Atlantic Biologicals, Miami, FL). HCC1937 and HCC1806 were grown in RPMI supplemented with 10% FBS. Cells were maintained in a humidified 37 °C incubator with 5% carbon dioxide.

Lee K.J., Wright G., Bryant H., Wiggins L.A., Schuler M, & Gassman N.R. (2020). EGFR signaling promotes resistance to CHK1 inhibitor prexasertib in triple negative breast cancer. Cancer Drug Resistance, 3(4), 980-991.

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Breast Cancer Cell Line Cultivation

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MCF10A cells were derived from benign proliferative breast tissue and are the most commonly used cell line to study normal breast cells.^{9 (link)} MDA-MB-231 cell line was isolated from a pleural effusion of a patient with invasive ductal carcinoma. This cell line is used to model late-stage breast cancer.^10,11 HCC1937 cell line (Hamon Cancer Center) was established from a grade III infiltrating ductal primary breast tumor from a patient with a germ-line BRCA1 mutation. The primary tumor showed large vacuoles in many of the cells suggestive of secretory variant of infiltrating intraductal carcinoma. HCC1937 cell line is homozygous for a BRCA1 5382insC mutation.^{12 (link)} Each cell line was purchased from American Type Culture Collection (ATCC; CRL-2336, HTB-26, CRL-10317). The MCF10A cells were cultured in DMEM/F12 supplemented with 20 ng mL⁻¹ hEGF, 100 ng mL⁻¹ cholera toxin, 10 μg mL⁻¹ bovine insulin, 500 ng mL⁻¹ hydrocortisone, 5% horse serum, and 1% penicillin/streptomycin. MDA-MB-231 cells were cultured in DMEM and supplemented with 10% FBS and 1% penicillin/streptomycin. HCC1937 cells were cultured in RPMI supplemented with 10% FBS and 1% penicillin/streptomycin.

Chappell K., Manna K., Washam C.L., Graw S., Alkam D., Thompson M.D., Zafar M.K., Hazeslip L., Randolph C., Gies A., Bird J.T., Byrd A.K., Miah S, & Byrum S.D. (2021). Multi-omics data integration reveals correlated regulatory features of triple negative breast cancer. Molecular Omics, 17(5), 677-691.

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Breast Cancer Cell Line Characterization

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MCF10A cells were derived from benign proliferative breast tissue and are the most commonly used cell line to study normal breast cells (9 ). MDA-MB-231 cell line was isolated from a pleural effusion of a patient with invasive ductal carcinoma. This cell line is used to model late-stage breast cancer (10 ,11 ). HCC1937 cell line (Hamon Cancer Center) was established from a grade III infiltrating ductal primary breast tumor from a patient with a germ-line BRCA1 mutation. The primary tumor showed large vacuoles in many of the cells suggestive of secretory variant of infiltrating intraductal carcinoma. HCC1937 cell line is homozygous for a BRCA1 5382insC mutation (12 ). Each cell line was purchased from American Type Culture Collection (ATCC; CRL-2336, HTB-26, CRL-10317). The MCF10A cells were cultured in DMEM/F12 supplemented with 20 ng/mL hEGF, 100 ng/mL cholera toxin, 10 μg/mL bovine insulin, 500 ng/mL hydrocortisone, 5% horse serum, and 1% Penicillin/Streptomycin. MDA-MB-231 cells were cultured in DMEM and supplemented with 10% FBS and 1% Penicillin/Streptomycin. HCC1937 cells were cultured in RPMI supplemented with 10% FBS and 1% Penicillin/Streptomycin.

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