End1 e6e7

Manufactured by Thermo Fisher Scientific

Sourced in United States

The End1/E6E7 is a lab equipment product designed for cell culture applications. It provides a defined, serum-free medium for the culture of human endocervical and ectocervical cells. The product supports the growth and maintenance of these cell types in vitro.

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5 protocols using end1 e6e7

Cervical Cancer Cell Line Culturing

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Human cervical cancer cell lines HeLa, SiHa, C-33A and ME-180, as well as the human cervical epithelium cell line End1/E6E7 were purchased from American Type Culture Collection (Manassas, VA, USA). HeLa, SiHa and C-33A cells were grown in high glucose Dulbeco's modified Eagle medium and ME-180 cells were grown in RPMI 1640 medium (both Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (Invitrogen; Thermo Fisher Scientific, Inc.), 1% streptomycin-penicillin solution and maintained at 37°C in a 5% CO₂-humidified incubator. Cells were passaged every 2–3 days. End1/E6E7 cells were grown in Keratinocyte serum-free medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 0.1 ng/ml human recombinant epithelial growth factor, 0.05 mg/ml bovine pituitary extract (both Santa Cruz Biotechnology, Inc., Dallas, TX, USA), 1% streptomycin-penicillin solution and maintained at 37°C in a 5% CO₂-humidified incubator.

Zhang Y., Yang Y., Liu R., Meng Y., Tian G, & Cao Q. (2019). Downregulation of microRNA-425-5p suppresses cervical cancer tumorigenesis by targeting AIFM1. Experimental and Therapeutic Medicine, 17(5), 4032-4038.

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Cell Culture Conditions for Cervical Cell Lines

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The human endocervical epithelial End1/E6E7 cell line, the human cervical cancer HeLa, C33A and SiHa cell lines, and the cervical squamous cell carcinoma Ca-Ski cell line were purchased from BioVector NTCC, Inc. End1/E6E7 cells were cultured in keratinocyte serum-free medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 50 µg/ml bovine pituitary extract (Absin Bioscience, Inc.), 0.1 ng/ml recombinant epidermal growth factor (Absin Bioscience, Inc.), 0.4 mmol/l CaCl₂ and 1% antibiotics (penicillin, 100 U/ml; streptomycin, 100 mg/ml). HeLa and CaSki cervical cancer cell lines were cultured in RPMI-1640 (Procell Life Science & Technology Co., Ltd.) containing 10% fetal bovine serum (Invitrogen; Thermo Fisher Scientific, Inc.) and 1% penicillin-streptomycin. C33A and SiHa cells were cultured in DMEM (MilliporeSigma) containing 10% fetal bovine serum (Invitrogen; Thermo Fisher Scientific, Inc.) and 1% penicillin-streptomycin. All cells were cultured under the conditions of 37°C with 5% CO₂.

Gao X, & Yang L. (2023). HBXIP knockdown inhibits FHL2 to promote cycle arrest and suppress cervical cancer cell proliferation, invasion and migration. Oncology Letters, 25(5), 186.

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Cervical Cell Lines for Research

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Cervical cancer cell lines HeLa and SiHa (ATCC, Manassas, VA, USA), and End1/E6E7, a human normal cervical epithelium cell line (ATCC), were obtained and employed for the cellular experiments in this study. End1/E6E7 cells were maintained in keratinocyte serum-free medium (K-SFM; Gibco, New York, NY, USA) containing 0.1 ng/ml epithelial growth factor (EGF; Gibco), 0.05 mg/ml bovine pituitary extract (Gibco), and 1% streptomycin–penicillin (SP; Invitrogen, Carlsbad, CA, USA) as previously reported1 . HeLa and SiHa cell lines were incubated in Dulbecco’s modified Eagle’s medium (DMEM; Gibco) containing 10% fetal bovine serum (FBS) and 1% SP at 37°C in 5% CO₂.

Yang M., Zhai X., Ge T., Yang C, & Lou G. (2018). miR-181a-5p Promotes Proliferation and Invasion and Inhibits Apoptosis of Cervical Cancer Cells via Regulating Inositol Polyphosphate-5-Phosphatase A (INPP5A). Oncology Research, 26(5), 703-712.

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Cell Culture Protocols for Diverse Cell Lines

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The LNcap cell line was purchased from American Type Culture Collection (Rockville, MD). LNcap cells and CEMX174 cells were maintained in cRPMI [RPMI 1640 (Gibco-BRL/Life Technologies, Gaithersburg, MD) supplemented with L-glutamine, 10 mM HEPES (pH 7.2), and 10% fetal bovine serum (FBS) (Hyclone, Logan, UT)]. CD4⁺ T cells were isolated from PBMC by negative selection with magnetic beads (Miltenyi Biotec) and were cultured in RPMI supplemented with 10% FCS (HyClone), 100 µg/mL streptomycin, and 100 U/mL penicillin. CD4⁺ T cells were activated with phytohemagglutinin (PHA) (3 ug/ml) in the presence of interleukin-2 (IL-2) (20 U/mL; Roche Applied Science) for 72 hrs. Human vaginal epithelial cell line VK2 (ATCC2616), ectocervical epithelial cell line Ect1/E6E7 (ATCCCRL1614) and endocervical cell line End1/E6E7 (ATCCCRL2615) were obtained from the American Type Culture Collection. These cells were selected because they were generated from normal vaginal, ectocervical and endocervical mucosal tissues; VK-2 cells and Ect1/E6E7 cells are derived from squamous epithelial cells lining the vagina and ectocervix respectively. End1/E6E7 cells are derived from simple columnar endocervical epithelium cells [35] (link). VK2, Ect1/E6E7 and End1/E6E7 cells were maintained in keratinocyte serum-free complete media (KSFM) (Gibco, Grand Island, NY).

Tang Y., George A., Nouvet F., Sweet S., Emeagwali N., Taylor H.E., Simmons G, & Hildreth J.E. (2014). Infection of Female Primary Lower Genital Tract Epithelial Cells after Natural Pseudotyping of HIV-1: Possible Implications for Sexual Transmission of HIV-1. PLoS ONE, 9(7), e101367.

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Cervical Cancer Cell Line Characterization

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Cervical cancer cell lines (SiHa, CaSki, ME180, SW756, C33A and HT3) were obtained from the American Type Culture Collection (ATCC) and maintained in IMDM media (Life Technologies) with 10% heat-inactivated FBS. Normal cervix epithelial cell lines Ect1/E6E7 and End1E6/E7 maintained in Keratinocyte-Serum Free medium (GIBCO-BRL 17005–042) with 0.1 ng/ml human recombinant EGF and 0.05 mg/ml bovine pituitary extract. HCvEpC were obtained from Cell Applications, Inc and maintained in HCvEpC media. Experiments were performed on cell lines under passage 30. Short Tandem Repeat (STR) profiling was last performed November 2020 which confirmed positive match and full cell line authentication per ATCC reference standards. Mycoplasma testing was performed every 3 months to verify no contamination. Sodium oleate, linoleic acid, protease inhibitor, phosphatase inhibitor cocktails, PFTα, GW9662 and lipid mixture (L0288) were purchased from Sigma. Arachidonic acid was purchased from Cayman chemicals. Adipocyte conditioned media was purchased from ZenBio Inc (CMA-10).

Muhammad N., Ruiz F., Stanley J., Rashmi R., Cho K., Jayachandran K., Zahner M.C., Huang Y., Zhang J., Markovina S., Patti G.J, & Schwarz J.K. (2022). Mono- and diunsaturated fatty acids sensitize cervical cancer to radiation therapy. Cancer research, 82(24), 4515-4527.

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