Human esophageal epithelial cell (HEEC), Eca109, and TE1 cells were purchased from the Shanghai Cell Bank (Shanghai, China). Cells were cultured in DMEM (Gibco, CA, USA) supplemented with 10% fetal bovine serum (Gibco-BRL) and antibiotics (100 U/mL penicillin and 100 U/mL streptomycin, HyClone Laboratories, Inc., USA).
Eca109
Manufactured by Shanghai Cell Bank
Sourced in China, United States
The Eca109 is a laboratory equipment designed for cell culture and analysis. It is a high-performance incubator that provides a controlled environment for cell growth and maintenance. The Eca109 features precise temperature, humidity, and gas mixture regulation to support a wide range of cell lines. It is a versatile and reliable tool for researchers and scientists working in the field of cell biology.
Automatically generated - may contain errors
Lab products found in correlation
Kyse150, by Shanghai Cell Bank (7 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Penicillin, by Cytiva (4 mentions)
Streptomycin, by Cytiva (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions)
Eca 109, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Epidermal growth factor (egf), by Merck Group (1 mentions)
Fibroblast growth factor (fgf), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Lonsera (1 mentions)
Lipofectamine 8000, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Solarbio (1 mentions)
Streptomycin, by Solarbio (1 mentions)
Penicillin streptomycin, by Beyotime (1 mentions)
Fetal bovine serum (fbs), by Beyotime (1 mentions)
Phosphate buffered saline pbs, by Beyotime (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
16 protocols using eca109
1
Culturing Human Esophageal Cell Lines
2
Cell Line Cultivation and Spheroid Culture
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Eca109, TE1 and HEEC cells were obtained from the Shanghai Cell Bank (Shanghai, China). After measured by mycoplasma detection, DNA-Fingerprinting, isozyme detection and cell vitality detection, these cell lines were immediately expanded and frozen such that they could be restarted every 3–4 months. All cell lines maintained in Dulbecco’s modified Eagle’s medium (Gibco, CA, USA) supplemented with 10% heat-inactivated foetal bovine serum (Gibco-BRL) and antibiotics (100 U/ml penicillin and 100 U/ml streptomycin; HyClone Laboratories, Inc., USA) at 37 °C in a humidified atmosphere of 5% CO2. Spheroids were cultured in F12/DMEM (Gibco, CA, USA) supplemented with EGF (Sigma, St Louis, USA), FGF (Gibco, CA, USA) and ITS (Sigma, St Louis, USA).
3
PIAS3 Knockdown in ESCC Cell Line
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Human ESCC cell line ECA109 (CC-Y1150) was purchased from Shanghai Cell Bank (Shanghai Biological Sciences, Chinese Academy of Sciences, Shanghai, China). The ECA109 cells were adherent cells and were cultured in Petri dishes. After successful resuscitation, the ECA109 cells were added to a complete culture medium composed of Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Carlsbad, CA) containing 100 U/mL of penicillin and 100 µg/mL of streptomycin (Solarbio, Beijing, China) and fetal bovine serum (FBS) (Lonsera, Shanghai, China) at a ratio of 9:1, and put into a 5% carbon dioxide (CO2), 37 ℃ incubator for routine culture, and the solution was changed the next day.
The small-interfering RNAs (siRNAs) targeting human PIAS3 were purchased from Genepharma (Shanghai, China) with the following sequences: PIAS3 siRNA-1#, 5'-GCAGGAACCCTTCTACAAA-3'; PIAS3 siRNA-2#, 5'-GGAGATCCATCAGAGAATA-3'; PIAS3 siRNA-3#, 5'-GTGATGAGATCCAATTCAT-3'. The ECA109 cells were transfected with 80 nM of siRNA using Lipofectamine 8000 (Invitrogen, CA, USA) for 48 h.
The small-interfering RNAs (siRNAs) targeting human PIAS3 were purchased from Genepharma (Shanghai, China) with the following sequences: PIAS3 siRNA-1#, 5'-GCAGGAACCCTTCTACAAA-3'; PIAS3 siRNA-2#, 5'-GGAGATCCATCAGAGAATA-3'; PIAS3 siRNA-3#, 5'-GTGATGAGATCCAATTCAT-3'. The ECA109 cells were transfected with 80 nM of siRNA using Lipofectamine 8000 (Invitrogen, CA, USA) for 48 h.
4
Culturing ESCC Cell Lines with F. nucleatum
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Human ESCC cell lines KYSE-150 and ECA-109 obtained from Shanghai Cell Bank, Chinese Academy of Sciences were cultured in RPMI-1640 medium (Gibco, Gaithersburg, MD, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Gaithersburg, MD, USA) and 1% penicillin/streptomycin (Beyotime, Shanghai, China) at 37°C in a humidified 5% CO2 atmosphere. The F. nucleatum strain ATCC 25,586 was purchased from the China General Microbiological Culture Collection Center (Beijing, China) and cultured in fastidious anaerobe broth (Tuopu biotechnology, Qingdao, China) at 37°C under anaerobic condition. For in vitro co-culture experiments, cells were washed with phosphate-buffered saline (PBS, Beyotime, Shanghai, China) and then incubated with F. nucleatum at a multiplicity of infection (MOI) of 100 in antibiotic-free RPMI-1640 medium supplemented with 10% FBS at 37°C under 5% CO2 for 24 h.
5
ESCC Cell Line Establishment and Culture
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Human ESCC tissues and adjacent tissues used in this study were obtained from Nanjing Medical University Affiliated Suzhou Hospital (Jiangsu, China). The tissue samples were immediately snap-frozen and stored at -80°C. All of the samples were obtained with informed consent and the study was approved by the Institutional Ethics Committee of Nanjing Medical University. Human ESCC cell lines, including ECA-109 and KYSE-150, as well as human esophageal epithelial cell line HEEC-1, were obtained from Shanghai Cell Bank (Shanghai, China). ECA-109 cells were cultured in DMEM medium, KYSE-150 cells and HEEC-1 cells were cultured in RPMI-1640 medium. All of the media (Hyclone, Logan, UT, USA) were supplemented with 10% FBS (Hyclone). The cells were incubated in a humidified atmosphere, with 5% CO2 at 37°C.
6
ESCC Tissue Acquisition and Cell Culture
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Human ESCC tissues and adjacent tissues used in this study were obtained from Nanjing Medical University Affiliated Suzhou Hospital (Jiangsu, China). The tissue samples were immediately snap-frozen and stored at −80°C for real-time PCR analysis and histological examination. All of the samples were obtained with informed consent and the study was approved by the Institutional Ethics Committee of Nanjing Medical University. Human ESCC cell lines, including ECA-109, TE-1, TE-10 and KYSE-150 were obtained from the Shanghai Cell Bank (Shanghai, China). ECA-109, TE-1 and TE-10 cells were cultured in DMEM medium, and KYSE-150 cells were cultured in RPMI-1640 medium. All of the media (Hyclone, USA) were supplemented with 10% FBS (Hyclone, USA). The cells were incubated in a humidified atmosphere, with 5% CO2 at 37°C.
7
Esophageal Squamous Cell Carcinoma Tissue Collection
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Tissues used in this study were collected from 44 patients with ESCC underwent esophagectomy at the First Affiliated Hospital of Nanjing Medical University. All collected tissue specimens were immersed in RNA Later stabilization solution (Qiagen) and were immediately frozen in liquid nitrogen and conserved at - 80 °C until RNA extraction. This study was approved by the Research Ethics Committee of the First Affiliated Hospital of Nanjing Medical University, and written informed consent was acquired from all the patients.
ESCC cell lines (Eca-109, Kyse-30, TE-1, Kyse-70, Kyse-150) and human esophageal mucosal epithelial cell line (Het-1A) were purchased from Shanghai Cell Bank (Shanghai, China). These cells were cultivated in RPMI-1640 or DMEM medium (GIBCO‐BRL) with 10% fetal bovine serum (FBS), 100 U/ml penicillin and 100 mg/ml streptomycin. Cells were incubated at 37 °C 5% CO2.
ESCC cell lines (Eca-109, Kyse-30, TE-1, Kyse-70, Kyse-150) and human esophageal mucosal epithelial cell line (Het-1A) were purchased from Shanghai Cell Bank (Shanghai, China). These cells were cultivated in RPMI-1640 or DMEM medium (GIBCO‐BRL) with 10% fetal bovine serum (FBS), 100 U/ml penicillin and 100 mg/ml streptomycin. Cells were incubated at 37 °C 5% CO2.
8
Cultivation of Esophageal Cell Lines
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The human Esophageal Epithelial Cells (HEEC), Eca109 and TE1 cells were purchased from the Shanghai Cell Bank (Shanghai, China). All cell lines cultured in Dulbecco's modified Eagle's medium (Gibco, CA, USA) supplemented with 10% heat-inactivated foetal bovine serum (Gibco-BRL) and antibiotics (100 U/ml penicillin and 100 U/ml streptomycin; HyClone Laboratories, Inc., USA).
9
Establishment of radioresistant esophageal cancer cell line
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The human ESC cell lines Eca109 were purchased from the Shanghai Cell Bank (Chinese Academy of Sciences) and cultured in RPMI1640 medium (Gibco, Waltham, USA) supplemented with 10% fetal bovine serum (Gibco, Waltham, USA) at 37°C in a 5% CO2 atmosphere.
The radioresistant cell line Eca109R was established by exposing Eca109 cells to 25 doses of 2Gy irradiation (total dose of 50 Gy), as previously reported [22 (link)]. To establish twist knockdown cells, four shRNA targeting twist mRNA and one negative control were designed (Supplementary Table 1 ), synthesized, and cloned separately into pYr-3.1 vector previously digested with BbsI and BamHI endonucleases. The pYr-3.1-shRNA plasmids were transfected into Eca109 or Eca109R cells with Lipofectamine 3000 according to the manufacturer's instructions (Thermo Fisher Scientific, NY). The cells were cultured in G418 (800 μg/ml) to screen for stable transfectants, and the selected twist-silenced or control cells were cultured long-term in medium supplemented with 400 μg/ml G418.
The radioresistant cell line Eca109R was established by exposing Eca109 cells to 25 doses of 2Gy irradiation (total dose of 50 Gy), as previously reported [22 (link)]. To establish twist knockdown cells, four shRNA targeting twist mRNA and one negative control were designed (
10
Cell Line Cultivation and Authentication
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All human ESCC cell lines including ECA‐109, TE‐1, KYSE‐150, and the normal human esophageal epithelial cell line (HEEC) were obtained from Shanghai Cell Bank (Shanghai Biological Sciences, Chinese Academy of Sciences, Shanghai, China). HEK293 cells were purchased from ATCC (Manassas, VA, USA). All cell lines were authenticated by morphological observation under a microscope and tested using short tandem repeat profiling. The cells were cultured in Dulbecco's modified Eagle's medium supplemented with 10% FBS, 1% penicillin/streptomycin, and incubated at 37 °C in a humidified atmosphere with 5% CO2.
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