Human ESCC cell lines Eca109 and TE-1 were purchased from Shanghai Institute of Biochemistry and Cell Biology (Shanghai, China). One human immortalized normal esophageal epithelial cell line (Het-1A), which was used as a ‘normal’ control for ESCC cell lines, was maintained in our laboratory. Eca109 and TE-1 cells were cultured in RPMI1640 (Hyclone) supplemented with 10% fetal bovine serum (Hyclone), 100 U/ml penicillin and 100 μg/ml streptomycin, within a humidified atmosphere containing 5% CO2 at 37°C. Het-1A cells were cultured in bronchial epithelial basal medium with growth supplements (Clonetics).
Growth supplements
Manufactured by Cambrex
Sourced in United States
Growth supplements are laboratory products designed to support the growth and development of various biological samples or cultures. These supplements provide essential nutrients, growth factors, and other compounds required for optimal growth and proliferation of cells, tissues, or microorganisms. The core function of growth supplements is to create an environment that promotes the healthy and consistent expansion of the target biological material.
Automatically generated - may contain errors
Lab products found in correlation
Ebm 2 basal media, by Cambrex (5 mentions)
Het 1a, by American Type Culture Collection (3 mentions)
Primary huvecs, by Cambrex (3 mentions)
Het 1a, by Cambrex (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Eca109, by Shanghai Institute of Biochemistry and Cell Biology (1 mentions)
Epilife, by Thermo Fisher Scientific (1 mentions)
Dksfm, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Primary human umbilical vein endothelial cells huvecs, by Cambrex (1 mentions)
9 protocols using growth supplements
1
Culturing Human ESCC Cell Lines
2
Esophageal Squamous Cell Cancer Tissue Collection
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Thirty pairs of primary esophageal squamous cell cancer tissues and corresponding adjacent normal esophageal tissues were obtained from untreated patients in Beijing Friendship Hospital (Capital Medical University) from 2009 to 2011 with informed consent and agreement. All tissue samples were snap frozen in liquid nitrogen and stored at −80°C until the extraction of RNA.
The human ESCC cell lines KYSE30, KYSE70, KYSE150, KYSE410, KYSE450, KYSE510, EC9706, and EC109 were kindly provided by the Cancer Institute and Hospital, Chinese Academy of Medical Science. The ESCC cell lines TE-1, TE-9, and TE-11 were gifts from Hebei Cancer Hospital of China. Het-1A, a human esophageal immortalized cell line, was purchased from the American Type Culture Collection. Five-week-old male BALB/c nu/nu mice were purchased from the Institute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences. ESCC cell lines used in this study are listed in previous reports [14 (link)-15 (link), 88 (link)-90 (link)].
All cell lines, except for Het-1A, were cultured in RPMI-1640 medium (Hyclone, USA) containing 10% fetal bovine serum (Gibco, USA) and 10 units / ml penicillin and streptomycin (Hyclone, USA). Cells were maintained at 37°C, 95% humidity, and 5% CO2. Het-1A cells were cultured in bronchial epithelial basal medium with growth supplements (Clonetics, USA).
The human ESCC cell lines KYSE30, KYSE70, KYSE150, KYSE410, KYSE450, KYSE510, EC9706, and EC109 were kindly provided by the Cancer Institute and Hospital, Chinese Academy of Medical Science. The ESCC cell lines TE-1, TE-9, and TE-11 were gifts from Hebei Cancer Hospital of China. Het-1A, a human esophageal immortalized cell line, was purchased from the American Type Culture Collection. Five-week-old male BALB/c nu/nu mice were purchased from the Institute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences. ESCC cell lines used in this study are listed in previous reports [14 (link)-15 (link), 88 (link)-90 (link)].
All cell lines, except for Het-1A, were cultured in RPMI-1640 medium (Hyclone, USA) containing 10% fetal bovine serum (Gibco, USA) and 10 units / ml penicillin and streptomycin (Hyclone, USA). Cells were maintained at 37°C, 95% humidity, and 5% CO2. Het-1A cells were cultured in bronchial epithelial basal medium with growth supplements (Clonetics, USA).
3
Immortalized Esophageal Cell Lines
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Immortalized human normal esophageal epithelial cell line Het-1A was purchased from ATCC (Manassas, VA). Immortalized esophageal epithelial NE-2 cells were gifted from Dr. Enmin Li. ESCC cell lines (KYSE series)56 (link) were provided by Dr. Y. Shimada. ZEC014 and ZEC134 cells were gifted from Dr. Dan Su. Het-1A cells were cultured in bronchial epithelial basal medium with growth supplements (Clonetics, USA). NE-2 cells were cultured in a 1:1 mixture of EpiLife and dKSFM (Gibco). KYSE series cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS) and antibiotics. And ZEC014 and ZEC134 cells were cultured in DMEM-F12 medium containing 10% FBS, 1% nonessential amino acid (NEAA) and antibiotics. All cell lines were maintained at 37 °C in humidified atmosphere containing 5% CO2. All of the cells were authenticated by short tandem repeat (STR) analysis and regularly tested for mycoplasma contamination.
4
Investigating Beta-adrenoceptor Modulation in ESCC
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The human ESCC cell lines KYSE410 and KYSE70 was obtained from the Chinese Academy of Medical Sciences and cultured in RPMI 1640 medium (Life Technologies, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (Life Technologies). The immortalized esophageal epithelial cell line HET-1A was purchased from the American Type Culture Collection (Manassas, VA, USA) and cultured in bronchial epithelium basal medium with growth supplements (Clonetics, San Diego, CA, USA).
Cells were plated at a density of 4×105 cells/well in 6-well plates. After 24 h of culture, the cells were treated for various purposes. To investigate the effect of beta-adrenoceptors, the cells were transfected with siRNA using Lipofectamine 2000 (Life Technologies) according to the manufacturer’s protocol [35 (link)]. siRNA sequences were as follows: beta1-adrenoceptor, 5′-CGCUCACCAACCUCUUCAUTT-3′ and 5′-AUGAAGAGGUUGGUGAGCGTT-3′; beta2-adrenoceptor, 5′-GCCUAGCGAUAACAUUGAUTT-3′and 5′-AUCAAUGUUAUCGCUAGGCTT-3′; negative control, 5′-UUCUCCGAACGUGUCACGUTT-3′ and 5′-ACGUGACACGUUCGGAGAATT-3′. To examine the effects of NNK, 0.1 μM NNK was added to the medium directly or at 6 h post-transfection according to our previous study (unpublished data). Cells were collected for further experimentation at 72 h post-treatment as described below.
Cells were plated at a density of 4×105 cells/well in 6-well plates. After 24 h of culture, the cells were treated for various purposes. To investigate the effect of beta-adrenoceptors, the cells were transfected with siRNA using Lipofectamine 2000 (Life Technologies) according to the manufacturer’s protocol [35 (link)]. siRNA sequences were as follows: beta1-adrenoceptor, 5′-CGCUCACCAACCUCUUCAUTT-3′ and 5′-AUGAAGAGGUUGGUGAGCGTT-3′; beta2-adrenoceptor, 5′-GCCUAGCGAUAACAUUGAUTT-3′and 5′-AUCAAUGUUAUCGCUAGGCTT-3′; negative control, 5′-UUCUCCGAACGUGUCACGUTT-3′ and 5′-ACGUGACACGUUCGGAGAATT-3′. To examine the effects of NNK, 0.1 μM NNK was added to the medium directly or at 6 h post-transfection according to our previous study (unpublished data). Cells were collected for further experimentation at 72 h post-treatment as described below.
5
Culturing Primary Human Endothelial Cells
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Primary HUVECs were obtained from Cambrex Bio Science (Charles City, IA) and were maintained using a previously described method (27 (link),28) (link). Briefly, cells were cultured until confluent at 37℃ at 5% CO2 in EBM-2 basal media supplemented with growth supplements (Cambrex Bio Science).
6
Culturing Primary Human Umbilical Vein Endothelial Cells
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Primary human umbilical vein endothelial cells (HUVECs) were obtained from Cambrex Bio Science (Charles City, IA, USA) and were maintained as described previously45 (link)46 (link)47 (link)48 (link). Briefly, cells were cultured in EBM-2 basal media supplemented with growth supplements (Cambrex Bio Science, Charles City, IA, USA) at 37 °C under 5% CO2 atmosphere until confluent. All experiments were performed with HUVECs at passage 3–5.
7
HUVEC Culture and Maintenance
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Primary human umbilical vein endothelial cells (HUVECs) were obtained from Cambrex Bio Science (Charles City, IA, USA) and were maintained as previously described57 (link)–59 (link). Briefly, cells were cultured to confluence in EBM-2 basal media supplemented with growth supplements (Cambrex Bio Science, Charles City, IA, USA) at 37 °C under an atmosphere of 5% CO2. All experiments were performed using HUVECs at passage 3–5.
8
Culturing Primary Human Umbilical Vein Endothelial Cells
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Primary HUVECs were obtained from Cambrex Bio Science (Charles City, IA, USA) and were maintained as described previously 16 . Briefly, cells were cultured in EBM‐2 basal media supplemented with growth supplements (Cambrex Bio Science, Charles City, IA, USA) at 37°C under 5% CO2 atmosphere until confluent. All experiments were performed with HUVECs at passage 3–5.
9
HUVEC Cell Culture Protocol
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Primary HUVECs were obtained from Cambrex Bio Science (Charles City, IA, USA) and were maintained as described previously41 (link), 42 (link). Briefly, cells were cultured in EBM-2 basal media supplemented with growth supplements (Cambrex Bio Science, Charles City, IA, USA) at 37 °C under a 5% CO2 atmosphere until confluent. All experiments were performed with HUVECs at passage 3–5.
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