ESCC lines (KYSE150, TE-1, TE-10, EC-1, and EC-109) and normal esophageal epithelial cells (NE1) were purchased from National Collection of Authenticated Cell Cultures (Shanghai, China). All cells were cultured in Dulbecco’s modified Eagle medium DMEM (Gibco, USA) or RPMI1640 media (Gibco) supplementary with 10% fetal bovine serum (FBS, Gibco), 100 U/ml penicillin and 100 μg/ml streptomycinn (both from SigmaAldrich, St. Louis, MO, USA)., with a humid incubator at 37 °C and 5% CO2.
Kyse150
Manufactured by National Collection of Authenticated Cell Cultures
Sourced in China, United States
The KYSE150 is a cell line that originated from a human esophageal squamous cell carcinoma. It is a commonly used model for esophageal cancer research.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (38 mentions)
Eca109, by National Collection of Authenticated Cell Cultures (34 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (21 mentions)
Kyse450, by National Collection of Authenticated Cell Cultures (19 mentions)
Ec9706, by National Collection of Authenticated Cell Cultures (18 mentions)
Kyse30, by National Collection of Authenticated Cell Cultures (17 mentions)
Het 1a, by National Collection of Authenticated Cell Cultures (16 mentions)
Ec109, by National Collection of Authenticated Cell Cultures (13 mentions)
Rpmi 1640, by Thermo Fisher Scientific (10 mentions)
Het 1a, by American Type Culture Collection (9 mentions)
Kyse510, by National Collection of Authenticated Cell Cultures (9 mentions)
Kyse 150, by Thermo Fisher Scientific (8 mentions)
Kyse140, by National Collection of Authenticated Cell Cultures (8 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (8 mentions)
Kyse410, by National Collection of Authenticated Cell Cultures (6 mentions)
Penicillin, by Thermo Fisher Scientific (6 mentions)
Streptomycin, by Thermo Fisher Scientific (6 mentions)
Te 10, by National Collection of Authenticated Cell Cultures (4 mentions)
Het 1a, by Lonza (4 mentions)
Eca9706, by National Collection of Authenticated Cell Cultures (4 mentions)
83 protocols using kyse150
1
Esophageal Cell Line Culture
2
Esophageal Squamous Cell Carcinoma Cell Lines
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ESCC cell lines including KYSE150, TE-1 (National Collection of Authenticated Cell Cultures, ShangHai, China), KYSE510, TE-7 (Guangzhou Jennio Biotech Co. Ltd., Guangzhou, China), and EC109 (National Infrastructure of Cell Line Resource, Beijing, China). Immortalized human esophageal squamous epithelial cell line Het-1A was purchased from ATCC (American Type Culture Collection, Manassas, VA, USA; Cat# CRL-2692TM). 293T cell line was purchased from (National Collection of Authenticated Cell Cultures). ESCC and 293T cell lines were cultured in RPMI-1640 and DMEM medium (HyClone™, USA), respectively, supplemented with 10% fetal bovine serum (FBS; GibcoTM, Thermo Scientific™, USA; Cat# 10099141C), 100 μg/ml streptomycin, and 100 units/ml penicillin (HyClone™). Het-1A cells were grown in BEGM™ Growth Medium (Lonza Walkersville Inc., USA). All cell lines were authenticated by STR and tested for mycoplasma recently.
3
Investigating miR-30d Inhibitor Effects on Human Cell Lines
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The human cell lines EC109, KYSE150, TE-1, TE-13, and HUVEC were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China) and cultured in DMEM medium (Gibco, USA) containing 10% fetal bovine serum. Ricolinostat (ACY-1215) was purchased from Selleck Chemicals (TX, USA) and was dissolved in DMSO to obtain a stock concentration of 100 mM. MiR-30d inhibitor was purchased from RiboBio Company (Guangzhou, China) and was dissolved in RNase-free water to obtain a concentration of 20 µM.
4
Culturing ESCC and Normal Esophageal Cells
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Human ESCC cell lines (Eca-109, EC-9706, KYSE-30, KYSE-150, TE-1) were purchased from Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Human normal esophageal epithelial cell line Het-1A was obtained from American Type Culture Collection (ATCC; Manassas, VA, USA). ESCC cells were cultured in RPMI 1640 medium (Invitrogen, Carlsbad, CA, USA) containing 10% fetal bovine serum and 1% penicillin/streptomycin. Het-1A cells were maintained in bronchial epithelial cell growth medium (BEGM, BulletKit, Lonza, MD). All cells were kept in a humidified incubator under an atmosphere of 5% CO2 at 37 °C.
5
Culturing and TGF-β Stimulation of ESCC Cells
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The human ESCC cell lines KYSE150 and TE1 were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China), where STR profile authentication and mycoplasma contamination test were also performed. Tumor cells were cultured in DMEM with 10% FBS, 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a humidified cell incubator with 5% CO2. For TGF-β stimulation, fibroblasts were cultured to 70–80% confluence, starved for 24 h in serum-free culture medium, and then treated with 10 ng/mL TGF-β1 (Selleck Chemicals, Houston, TX, USA) for the indicated times.
6
Investigating LETM1 and KIF14 in Esophageal Cancer
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Human esophageal epithelial cell line (Het-1A) and human ESCC cell lines (KYSE150, TE10, HCE7, KYSE450 and TE11) were purchased from the Cell Bank of the Chinese Academy of Sciences. Cells were cultured in Dulbecco's modified eagle medium (DMEM; Gibco) containing 10% fetal bovine serum (FBS) with 5% CO2 at 37°C and passaged on alternate days. The cell lines with the highest expression of LETM1 or KIF14 were used for the subsequent experiments.
For transfection, small interfering RNA (siRNA) targeting LETM1 (siRNA-LETM1-1 and siRNA-LETM1-2), corresponding negative control (siRNA-NC), KIF14 overexpression plasmids (Ov-KIF14) and pcDNA3.1 empty vector (Ov-NC) were obtained from Shanghai GenePharma company and transfected into TE11 cells by Lipofectamine™ 2000 transfection reagent (Thermo Fisher Scientific, Inc.) according to the manufacturer’s instruction.
For transfection, small interfering RNA (siRNA) targeting LETM1 (siRNA-LETM1-1 and siRNA-LETM1-2), corresponding negative control (siRNA-NC), KIF14 overexpression plasmids (Ov-KIF14) and pcDNA3.1 empty vector (Ov-NC) were obtained from Shanghai GenePharma company and transfected into TE11 cells by Lipofectamine™ 2000 transfection reagent (Thermo Fisher Scientific, Inc.) according to the manufacturer’s instruction.
7
Esophageal cancer cell line culture
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The ESCC cell lines TE-1, ECA109 and KYSE-150, and normal esophageal squamous epithelium cell line Het-1a were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). All cells were cultured in RPMI-1640 medium containing 10% FBS, 100 IU/ml penicillin and streptomycin at 37°C, 5% CO2. All media were purchased from Gibco (Grand Island, NY, USA). The following siRNA sequences were designed and synthesized by GenePharma Company (GenePharma, Shanghai, China): let-7a mimics 5′-UGAGGUAGUAGGUUGUAUAGUU-3′ (forward), 5′-CUAUACAACCUACUACCUCAUU-3′ (reverse); negative control (NC) for mimics 5′-UUCUCCGAACGUGUCACGUTT-3′ (forward), 5′-ACGUGACACGUUCGGAGAATT-3′ (reverse); let-7a inhibitor 5′-AACUAUACAACCUACUACCUCA-3′; NC for inhibitor 5′-CAGUACUUUUGUGUAGUACAA-3′. Mimics (50 nM) or inhibitors (100 nM) were transfected into ECA-109 cell lines with Lipofectamine 2000 reagents (Invitrogen, Shanghai, China) according to the manufacturer's instructions. Untransfected groups were cultured under normal conditions.
8
Esophageal Cancer Cell Lines Analysis
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Human ESCC cell lines (Eca-109, KYSE-150 and TE-1) were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Human normal esophageal cell line (Het-1A) was purchased from ATCC (American Type Culture Collection, Manassas, USA). Cell culture, total RNA extraction and qRT-PCR analysis were carried out as described previously28 (link). The relative expression of each gene in prognostic signature was calculated after normalization to GAPDH. Primer sequences are listed in Supplementary Table 2 .
9
Culturing ESCC Cell Lines
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Human ESCC cell lines Kyse150, Eca109, Eca9706 and TE‐1 were purchased from the cell bank of the Chinese Academy of Sciences and cultured in RPMI‑1640 medium (Gibco; Thermo Fisher Scientific, Inc, Waltham, MA, USA) containing 10% foetal bovine serum (FBS, Gibco; Thermo Fisher Scientific, Inc), streptomycin (100 mg/ml) and penicillin (100 U/ml). All cells were cultured in a humidified incubator at 37℃ with 5% CO2.
10
Characterization of esophageal cancer cell lines
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Written informed consent was obtained from each patient before the study began, and all of the patients were followed up on a regular basis. The detailed clinicopathological characteristics are listed in Tables S1-2.
Cell culture and reagents HEK293T cells, human ESCC cell lines (ECA109, TE1, TE10, KYSE30, KYSE150, KYSE410 and EC9706), and human normal esophageal epithelial cell lines (HEEC and Het-1a) were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). HEK293T and HEEC cells were cultured in DMEM (Gibco, MA, USA) supplemented with 10% FBS (Gibco, MA, USA). ECA109, TE1, TE10, KYSE150 and EC9706 cells were cultured in RPMI-1640 medium (Gibco, MA, USA) supplemented with 10% FBS. KYSE30 and KYSE410 cells were cultured in F12-K medium (Gibco, MA, USA) supplemented with 10% FBS. Het-1a cells were cultured using BEGMTM BulletKit TM (Lonza, GA, USA). All cell lines were cultured at 37°C with 5% CO 2 , authenticated by the short tandem repeat (STR) method and evaluated for mycoplasma contamination.
Cell culture and reagents HEK293T cells, human ESCC cell lines (ECA109, TE1, TE10, KYSE30, KYSE150, KYSE410 and EC9706), and human normal esophageal epithelial cell lines (HEEC and Het-1a) were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). HEK293T and HEEC cells were cultured in DMEM (Gibco, MA, USA) supplemented with 10% FBS (Gibco, MA, USA). ECA109, TE1, TE10, KYSE150 and EC9706 cells were cultured in RPMI-1640 medium (Gibco, MA, USA) supplemented with 10% FBS. KYSE30 and KYSE410 cells were cultured in F12-K medium (Gibco, MA, USA) supplemented with 10% FBS. Het-1a cells were cultured using BEGMTM BulletKit TM (Lonza, GA, USA). All cell lines were cultured at 37°C with 5% CO 2 , authenticated by the short tandem repeat (STR) method and evaluated for mycoplasma contamination.
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