HCT-116 and LS-174T cell lines were obtained from ATCC (Manassas, VA) in 2014. Parental LS-174T (ATCC), LS-174T/vector, LS-174T/shPPARδ, LS-174T/shNanog, parental HCT-116 (ATCC), HCT-116/WT, HCT-116/PPARd−/−, HCT-116/vector, and HCT-116/shNanog cells were maintained in McCoy’s 5A medium (Life Technologies) with 10% fetal bovine serum (FBS) (Hyclone, Logan, UT). All CRC cells are used between passages 2 to 5. All cell lines have been tested by MycoProbe Mycoplasma Detection Kit (R&D) and also authenticated before the experiment according ATCC STR database. For GW501516 treatment, the cells were cultured with serum-free medium for 24 or 48 h and then treated with vehicle or indicated concentration of GW501516 for 24 h. After treatment, the cells were subjected to in vitro sphere-forming assay, subcutaneous (sub-Q) injection of NSG mice, q-PCR, Western blotting, luciferase assay, and ChIP assay.
Ls174t
Manufactured by Thermo Fisher Scientific
Sourced in United States, China
The LS174T is a cell line derived from a human colon adenocarcinoma. It is commonly used in cancer research and drug development studies.
Automatically generated - may contain errors
Lab products found in correlation
Hct116, by Thermo Fisher Scientific (26 mentions)
Ls174t, by American Type Culture Collection (26 mentions)
Sw480, by Thermo Fisher Scientific (23 mentions)
Hct116, by American Type Culture Collection (20 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (20 mentions)
Sw480, by American Type Culture Collection (20 mentions)
Sw620, by Thermo Fisher Scientific (16 mentions)
Sw620, by American Type Culture Collection (15 mentions)
Rpmi 1640, by Thermo Fisher Scientific (11 mentions)
Ht 29, by Thermo Fisher Scientific (11 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (11 mentions)
Hct15, by Thermo Fisher Scientific (10 mentions)
Hct15, by American Type Culture Collection (10 mentions)
Dld 1, by American Type Culture Collection (10 mentions)
Dld 1, by Thermo Fisher Scientific (9 mentions)
Dmem medium, by Thermo Fisher Scientific (8 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (8 mentions)
Caco 2, by Thermo Fisher Scientific (7 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (6 mentions)
Caco 2, by American Type Culture Collection (6 mentions)
43 protocols using ls174t
1
Colorectal Cancer Cell Lines Characterization
2
Comprehensive Colorectal Cancer Cell Line Profiling
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Human colon epithelial cell line NCM460 and human CRC cell lines Lovo, SW48, T84, SW948, LS174T, HT29, HCT116 and SW480 were purchased from Procell Life Sciences Co. Ltd., (Wuhan, Hubei, China). Cell lines NCM460 was cultured in RPMI-1640 (Gibco) medium, T84 in DMEM/F12 (Gibco) medium, Lovo in F12K (Gibco) medium, LS174T in MEM, HT29 and HCT116 in (Gibco) McCoy's 5A, SW48, SW948 and SW480 in L15 (Gibco) medium.
All media for the human cell lines were supplemented with 10% FBS and 1% penicillin/streptomycin antibiotics. SW48, SW948 and SW480 cell lines were cultured in 100% air at 37 °C. Other cell lines were cultured under 95% air-5% CO2 at 37 °C. All cell lines were validated by STR DNA finger-printing. Experiments were carried out within 6 months after acquisition of the cell lines. In addition, mycoplasma contamination was ruled out using a PCR-based method.
HT29 and LS174T cells were transfected with miR-20a mimic and negative control (NC) or miR-20a inhibitor and inhibitor NC (inNC) respectively using Lipofectamine 2000 (Invitrogen, USA). After 48 h transfection, these cells were harvested for further experiments. These inhibitors and activator, including C75 (inhibitor of FASN), PKI-402 (inhibitor of PI3K/mTOR), MSAB (inhibitor of Wnt/β-catenin), Mycro 3 (inhibitor of Myc) and ZLN024 (activator of AMPK), were purchased from GLPBIO.
All media for the human cell lines were supplemented with 10% FBS and 1% penicillin/streptomycin antibiotics. SW48, SW948 and SW480 cell lines were cultured in 100% air at 37 °C. Other cell lines were cultured under 95% air-5% CO2 at 37 °C. All cell lines were validated by STR DNA finger-printing. Experiments were carried out within 6 months after acquisition of the cell lines. In addition, mycoplasma contamination was ruled out using a PCR-based method.
HT29 and LS174T cells were transfected with miR-20a mimic and negative control (NC) or miR-20a inhibitor and inhibitor NC (inNC) respectively using Lipofectamine 2000 (Invitrogen, USA). After 48 h transfection, these cells were harvested for further experiments. These inhibitors and activator, including C75 (inhibitor of FASN), PKI-402 (inhibitor of PI3K/mTOR), MSAB (inhibitor of Wnt/β-catenin), Mycro 3 (inhibitor of Myc) and ZLN024 (activator of AMPK), were purchased from GLPBIO.
3
Cell Line Culture Conditions
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MKN45, KATOIII and LS174T were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). These cell lines were authenticated by DNA short tandem repeat profiling, and experiments were carried out within 6 months of resuscitation. Cell lines were routinely assessed by cell morphology and their average doubling time. All cell lines were maintained in a humidified 5% CO2 incubator at 37°C in their respective medium as follows: MKN45 in RPMI-1640 medium, KATOIII in IMDM, and LS174T in EMEM (Invitrogen, Carlsbad, CA, USA). The culture media used were all supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin, with the exception of IMDM being supplemented with 20% FBS. The culture medium for LS174T was further supplemented with 2 mM Glutamine and 1% Non-Essential Amino Acids (All from Invitrogen, Carlsbad, CA, USA).
4
Validation of Colorectal Cancer Cell Lines
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Human colon epithelial cell line NCM460 and human CRC cell lines Lovo, SW48, T84, SW948, LS174T, HT29, HCT116 and SW480 were purchased from Procell Life Sciences Co. Ltd., (Wuhan, Hubei, China).
Cell lines NCM460 was cultured in RPMI-1640 (Gibco) medium, T84 in DMEM/F12 (Gibco) medium, Lovo in F12K (Gibco) medium, LS174T in MEM, HT29 and HCT116 in (Gibco) McCoy's 5A, SW48, SW948 and SW480 in L15 (Gibco) medium.
All media for the human cell lines were supplemented with 10% FBS and 1% penicillin/streptomycin antibiotics. SW48, SW948 and SW480 cell lines were cultured in 100% air at 37 °C. Other cell lines were cultured under 95% air-5% CO 2 at 37 °C. All cell lines were validated by STR DNA nger-printing.
Experiments were carried out within 6 months after acquisition of the cell lines. In addition, mycoplasma contamination was ruled out using a PCR-based method.
HT29 and LS174T cells were transfected with miR-20a mimic and negative control (NC) or miR-20a inhibitor and inhibitor NC (inNC) respectively using Lipofectamine 2000 (Invitrogen, USA). After 48 h transfection, these cells were harvested for further experiments.
Cell lines NCM460 was cultured in RPMI-1640 (Gibco) medium, T84 in DMEM/F12 (Gibco) medium, Lovo in F12K (Gibco) medium, LS174T in MEM, HT29 and HCT116 in (Gibco) McCoy's 5A, SW48, SW948 and SW480 in L15 (Gibco) medium.
All media for the human cell lines were supplemented with 10% FBS and 1% penicillin/streptomycin antibiotics. SW48, SW948 and SW480 cell lines were cultured in 100% air at 37 °C. Other cell lines were cultured under 95% air-5% CO 2 at 37 °C. All cell lines were validated by STR DNA nger-printing.
Experiments were carried out within 6 months after acquisition of the cell lines. In addition, mycoplasma contamination was ruled out using a PCR-based method.
HT29 and LS174T cells were transfected with miR-20a mimic and negative control (NC) or miR-20a inhibitor and inhibitor NC (inNC) respectively using Lipofectamine 2000 (Invitrogen, USA). After 48 h transfection, these cells were harvested for further experiments.
5
Culturing Colorectal Cancer Cell Lines
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All cell lines (Caco-2, HT29, SW480, DLD1, HCT116, LS174T, RKO) were cultured in a humidified atmosphere (37°C, 5% CO2) in their corresponding medium supplemented with 10% fetal calf serum (Life Technologies) and 1% Glutamin (GE Healthcare). Caco-2, LS174T, RKO and SW480 were cultured in DMEM, high glucose, DLD1 in RPMI, HCT116 and HT29 in McCoys 5A medium (all Life Technologies). All CRC cell lines were screened for mycoplasma contamination using Venor GeM Classic Mycoplasma Detection Kit (Minerva biolabs) and DNA fingerprinting was performed for all seven cell lines at the Leibniz Institute DSMZ, Braunschweig, Germany.
6
Culturing Human Colorectal Cancer Cell Lines
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The human CRC cell lines (SW480, SW620, HCT15, HCT116, Ls174t, HT29, Caco-2, Colo205, KM-12 and DLD1) were initially purchased from American Type Culture Collection (Manassas, VA, USA). SW620 and HT29 were cultured in DMEM medium (Invitrogen, Carlsbad, CA, USA) with 10% FBS (Gibco). SW480, HCT15, HCT116, Ls174t, Caco-2, Colo205, KM-12 and DLD1 were cultured in RPMI 1640 medium (Invitrogen, Carlsbad, CA, USA) containing 10% FBS (Gibco).
7
Cultivation of Gastrointestinal Cancer Cell Lines
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Human gastric carcinoma cell lines MKN45 and KATO-III were obtained from the Cancer Research Campaign Laboratories (University of Nottingham, NG7 2RD, UK) and the American Type Culture Collection (ATCC, Manassas, VA, USA), respectively. HT29-5F12 and HT29-5M21 colon adenocarcinoma cells were a kind gift from Dr Thécla Lesuffleur (Université Pierre et Marie Curie, Paris, France). LS174T colon adenocarcinoma cell line was purchased from Sigma-Aldrich (Sigma-Aldrich, MO, USA). All cell lines were maintained in a humidified atmosphere of 95% air and 5% CO2 at 37°C in their respective media as follows: MKN45 in RPMI-1640 medium, KATO-III in Iscove’s Dulbecco’s modified Eagle’s medium, HT29-5F12 and HT29-5M21 in Dulbecco’s modified Eagle’s medium and LS174T in Minimum Essential Medium Eagle EBSS medium (all from Invitrogen, Carlsbad, CA, USA). The culture media used were all supplemented with 10% (v/v) fetal bovine serum and 1% (v/v) penicillin-streptomycin (Invitrogen, Carlsbad, CA, USA), with the exception of Iscove’s Dulbecco’s modified Eagle’s medium being supplemented with 20% fetal bovine serum. As per the distributor’s instructions, the culture medium for LS174T was further supplemented with 2 mM Glutamine and 1% Non-Essential Amino Acids (NEAA).
8
Colorectal Cancer Cell Lines and Molecular Markers
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The HT29, SW480, SW620, LS174T, HCT116, C2BBel and HEK293T cell lines were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA). HT29, SW480, HCT116 and C2BBel cells were maintained in RPMI-1640 medium (Invitrogen, Carlsbad, CA, USA) supplemented with 10% heat-inactivated foetal bovine serum (FBS, Invitrogen); LS174T and HEK293T were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen) supplemented with 10% FBS, and SW480 cells were maintained in L15 medium (Invitrogen) supplemented with 10% FBS. The cells were incubated at 37 °C with 5% CO2. The following commercial antibodies were obtained from Cell Signaling (Danvers, MA, USA): rabbit monoclonal to PKM2; phospho-STAT3 (Y705); cleaved PARP; cleaved caspase-3; cyclin D1; p27Kip1; and mouse monoclonal to STAT3. The secondary antibodies used in the experiments were IRDye® 680 donkey anti-mouse IgG, IRDye® 800CW goat anti-rabbit IgG (LI-COR Biosciences, Lincoln, NE, USA), and Alexa Fluor 594 goat anti-rabbit IgG (Invitrogen). Gefitinib and STAT3 inhibitor V (Stattic) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
9
Cell Culture Protocols for Cancer Research
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HEK293T, Huh7, RKO, U2OS, 3T3-L1, C3H10T1/2 clone 8, HCT116, Ls174t, SW480, HT-29, DLD1, and Caco-2 were purchased from American Type Culture Collection (ATCC), YAPC and ST2 were obtained from DSMZ. HEK293T, YAPC, Huh7, SW480 and 3T3-L1 were grown in DMEM (Cat#11995-040, Invitrogen) with 10% FBS (Cat#1500-500, Seradigm) and penicillin/streptomycin (Cat#10378016, Invitrogen). ST2 and DLD1 were maintained in RPMI-1640 (Cat#22400-071, Invitrogen) with 10% FBS and penicillin/streptomycin. U2OS, HT-29 and HCT116 were grown in McCoy’s 5A (Cat#16600-082, Invitrogen) with 10% FBS and penicillin/streptomycin. RKO, Caco-2, Ls174t and C3H10T1/2 were maintained in MEM (Cat#11095-080, Invitrogen) with 10% FBS and penicillin/streptomycin. All cultures were grown at 37 °C in a 5% CO2 incubator. Cell lines were tested to be free of mycoplasma contamination, and were not subjected to extra authentication steps.
Plasmids were generated using standard recombinant DNA techniques. Site-specific mutagenesis was performed using Q5 Site-Directed Mutagenesis Kit (New England Biolabs). Primers used for cloning are shown in Supplementary Table1 .
Plasmids were generated using standard recombinant DNA techniques. Site-specific mutagenesis was performed using Q5 Site-Directed Mutagenesis Kit (New England Biolabs). Primers used for cloning are shown in Supplementary Table
10
Culturing Colon Epithelial and Cancer Cells
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Normal human colon epithelial cells (CCD 841 CoN cells) and four CRC cell lines (SW480, LoVo, SW620 and LS174T) were obtained from Invitrogen; Thermo Fisher Scientific, Inc. The cells were cultured in RPMI 1640 medium (Gibco; Thermo Fisher Scientific, Inc.) containing 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.), 0.03% glutamine and 100 µg/ml streptomycin at 37°C in an incubator with 5% CO2. The cells were digested and subcultured with 0.25% trypsin when confluence reached 80–90%.
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