Normal human colon epithelial cell line (FHC) and five human CRC cell lines (HCT-116, HT29, LoVo, SW480, and SW620) were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA). FHC cells were cultured in DMEM/F12 medium (ATCC) containing 25 mM HEPES, 10 ng/ml cholera toxin, 5 μg/ml insulin and transferrin, and 100 ng/ml hydrocortisone. LoVo cells, HCT-116 and HT29 cells, and SW480 and SW620 cells were grown in F-12K medium, McCoy’s 5a medium modified, and Leibovitz’s L-15 medium (all ATCC), respectively. All of the media were supplemented with 10% fetal bovine serum (FBS; Gibco, Grand Island, NY, USA). Cells were maintained at 37°C in a humidified incubator under 5% CO2 condition10 (link).
Sw480
Manufactured by American Type Culture Collection
Sourced in United States, China, Germany, Japan, France, Poland, Australia, Israel, United Kingdom, Italy
The SW480 is a laboratory equipment product from the American Type Culture Collection (ATCC). It is a colorectal adenocarcinoma cell line that can be used for various cell-based research applications.
Automatically generated - may contain errors
Lab products found in correlation
Hct116, by American Type Culture Collection (1194 mentions)
Sw620, by American Type Culture Collection (811 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (593 mentions)
Dld 1, by American Type Culture Collection (392 mentions)
Caco 2, by American Type Culture Collection (307 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (258 mentions)
Ht 29, by American Type Culture Collection (206 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (181 mentions)
Sw480, by Thermo Fisher Scientific (176 mentions)
Ncm460, by American Type Culture Collection (165 mentions)
Hct15, by American Type Culture Collection (153 mentions)
Ls174t, by American Type Culture Collection (147 mentions)
Hct116, by Thermo Fisher Scientific (144 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (142 mentions)
Penicillin, by Thermo Fisher Scientific (132 mentions)
Streptomycin, by Thermo Fisher Scientific (130 mentions)
Rpmi 1640, by Thermo Fisher Scientific (122 mentions)
Sw620, by Thermo Fisher Scientific (94 mentions)
Hek293t, by American Type Culture Collection (93 mentions)
Colo205, by American Type Culture Collection (92 mentions)
1 811 protocols using sw480
1
Culturing of Human Colon Cell Lines
2
Culturing Colorectal Cancer Cell Lines
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All CRC cell lines (SW480, SW620, HT-29, COLO 205, and WiDr) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA), and cultured in Dulbecco's Modified Eagle Medium (DMEM; Hyclone, Logan, UT, USA) supplemented with 10% fetal bovine serum (FBS; Hyclone) and 10 U/mL of penicillin-streptomycin (Hyclone). All cell lines were routinely tested for mycoplasma contamination and cell lines used in subsequent experiments (SW480 and SW620) were authenticated by short tandem repeat profiling at the Research Institute of National Cancer Center (Republic of Korea).
HUVEC cells were purchased from ATCC, and cultured in Endothelial Cell Growth medium (EGM-2; Lonza, Walkersville, MD, USA) supplemented with 10% FBS.
HUVEC cells were purchased from ATCC, and cultured in Endothelial Cell Growth medium (EGM-2; Lonza, Walkersville, MD, USA) supplemented with 10% FBS.
3
Optimizing Cell Culture Conditions for In Vitro Studies
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The HCT116, HT-29, LOVO, SW480, and NCM460 cell lines were obtained from the American Type Culture Collection (ATCC), USA. To ensure proper growth and maintenance of phenotypic characteristics, each cell line was cultured under its specific optimal conditions as recommended by ATCC. HCT116 and HT-29 cells were cultured in McCoy’s 5 A Medium modified (ATCC, USA), supplemented with 10% fetal bovine serum (FBS), at 37 °C in a humidified atmosphere containing 5% CO2. LOVO cells were grown in F-12 K Medium (ATCC, USA) with 10% FBS, and NCM460 cells were cultured in M3:10™ Medium (INCELL Corporation, USA) specifically designed to support their normal phenotype, at 37 °C in an atmosphere of 5% CO2. SW480 cells were maintained in Leibovitz’s L-15 Medium (ATCC, USA) supplemented with 10% FBS, at 37 °C in a non-CO2 environment (100% Air). These specific culture conditions are critical to ensure that the cells express relevant phenotypes and that gene expression and transcription processes are not adversely affected.
4
Epigenetic Regulation of Colorectal Cancer
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Human normal colonic epithelial cell line FHC (ATCC® CRL-1831), as well as human CRC cell lines HCT116 (ATCC® CCL-247™, Rockville, MD) and SW480 (ATCC® CCL-228™), was procured from ATCC (Manassas, VA). HCT116 cells were cultured in McCoy’s 5a medium containing 10% FBS (ATCC, No. 30-2007), while SW480 cells were maintained in Leibovitz’s L-15 medium (ATCC, No. 30-2008) containing 10% FBS. Meanwhile, FHC cells were cultured in DMEM/F12 medium. All the aforementioned cells were cultured in a humid incubator at 37 °C with 5% CO2 in air.
Subsequently, the cells were seeded into a six-well plate (3 × 105 cells/well). Upon reaching 50% cell confluence, transfection was carried out using Lipofectamine 2000 reagent (11668-019, Invitrogen, Carlsbad, CA). siRNAs targeting SP1 (si-SP1-1, si-SP1-2), siRNAs targeting TUG1 (si-TUG1-1, si-TUG1-2), siRNA targeting KDM2A (si-KDM2A) and its negative control (si-NC), miR-421 mimic, mimic NC, TUG1 over-expression plasmid (oe-TUG1) and oe-NC were all synthesized and provided by GenePharma (Shanghai, China). The siRNA sequences are shown in Supplementary Table1 .
Subsequently, the cells were seeded into a six-well plate (3 × 105 cells/well). Upon reaching 50% cell confluence, transfection was carried out using Lipofectamine 2000 reagent (11668-019, Invitrogen, Carlsbad, CA). siRNAs targeting SP1 (si-SP1-1, si-SP1-2), siRNAs targeting TUG1 (si-TUG1-1, si-TUG1-2), siRNA targeting KDM2A (si-KDM2A) and its negative control (si-NC), miR-421 mimic, mimic NC, TUG1 over-expression plasmid (oe-TUG1) and oe-NC were all synthesized and provided by GenePharma (Shanghai, China). The siRNA sequences are shown in Supplementary Table
5
Characterization of Colorectal Cancer Cell Lines
6
Colon and Colorectal Cancer Cell Lines
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A normal human colon epithelial cell line, NCM460, was obtained from Lonza (Basel, Switzerland) and cultured in DMEM: F12 medium (Gibco, Waltham, MA, USA) supplemented with 10% FBS (Invitrogen, Waltham, MA, USA). Four human colorectal cancer cell lines, HCT116 (p53wt; CCL-247™), RKO (p53wt; CRL-2577™), LoVo (p53wt; CCL-229™), and SW480 (p53mut; CCL-228™), were procured from ATCC (Manassas, VA, USA). HCT116 cells were cultured in McCoy’s 5a medium modified (Gibco) supplemented with 10% FBS (Invitrogen, CA, USA). RKO cells were cultured in an Eagle’s Minimum Essential Medium (Gibco) supplemented with 10% FBS (Invitrogen). LoVo cells were cultured in F-12K medium (ATCC) supplemented with 10% FBS (Invitrogen). SW480 cells were cultured in Leibovitz’s L-15 medium (ATCC) supplemented with 10% FBS (Invitrogen). All cells were cultured at 37°C in 5% CO2.
7
Culturing Colorectal Cancer Cell Lines
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Colorectal cancer cell lines CaCo2, HCT116 and SW480 were acquired from ATCC (Manassas, VA, USA). All cells are adherent with epithelial morphology. CaCo2 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Biowest, Nuaillé, France) supplemented with 20% fetal bovine serum (FBS) (GE Healthcare-Hyclone, Chicago, IL, USA); HCT116 cells were cultured in McCoy’s 5A (Biowest, Nuaillé, France) supplemented with 10% FBS and SW480 cells in L-15 medium (ATCC) with 10% FBS. A mixture of antibiotics including 100 U/mL penicillin and 100 μg/mL streptomycin (Lonza, Basel, Switzerland) was added to all cell cultures. Cells were incubated at 37 °C and 5% CO2 in a humidified incubator (HeraCell, Heraeus, Hanau, Germany).
8
Human Colon Cancer Cell Line Culture and CD137L Signaling Induction
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The four human colon cancer cell lines HT-29, HCT-116, SW480, and SW620 were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). HT-29 and HCT-116 cells were cultured in McCoy Medium (ATCC) supplemented with 10% (v/v) fetal bovine serum (FBS) and 1% (v/v) penicillin/streptomycin (pen/strep). SW480 and SW620 cells were maintained in RPMI 1640 medium (ATCC) containing 10% (v/v) FBS and 1% (v/v) pen/strep. All cell lines were incubated at 37°C in 5% CO2. To induce CD137L signaling cell culture plates were coated with 10 µg/ml recombinant human CD137-Fc (R&D Systems, Minneapolis, MN, USA) at 4°C overnight. To preserve cell surface epitopes cells were then carefully detached using accutase solution (Sigma-Aldrich, St. Louis, MO, USA) and seeded in the pre-coated wells in a defined number.
9
Colorectal Cancer Cell Line Cultivation
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Human normal colorectal mucosal cell line (FHC) and human CRC cell lines (LOVO, HT-29, HCT116, SW480) in this study, were available from American Type Culture Collection (ATCC, Manassas, VA). FHC and LOVO cells were cultured in F-12K medium (Invitrogen, Carlsbad, CA, USA). HT-29 and HCT116 cells were cultured in ATCC-formulated McCoy's 5a medium (Manassas, VA). SW480 cell was cultured in ATCC-formulated Leibovitz's L-15 medium (Manassas, VA). All mediums with 10%FBS (Gibco) and 1% antibiotics were utilized for cell culture under the condition of 37 o C and 5% CO 2 . Besides, 3 U/mg of RNase R from Epicentre Technologies (Madison, WI) and 2 mg/ml of Actinomycin D from Sigma-Aldrich (St. Louis, MO) were used to treat cell samples of HCT116 and SW480.
10
Intestinal Epithelial Cell Lines: Cultivation and Stimulation
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The intestinal epithelial cell lines including HT29, SW620, SW480, LoVo, HCT116, Caco‐2 and IEC‐6 were purchased from American Type Culture Collection, and cultured routinely in RPMI 1640 (HT29, SW620, SW480, LoVo and HCT116) or DMEM (Caco‐2 and IEC‐6) medium respectively, supplemented with 10% or 20% FBS at 37°C in 5% CO2. Cultured or transfected cells were incubated with 1 μM PMA for 15 min. or 1 μg/ml LPS from E. coli for 24 hrs, then processed as follows.
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