Ls180

Manufactured by American Type Culture Collection

Sourced in United States

The LS180 is a laboratory centrifuge designed for general-purpose applications. It has a maximum speed of 18,000 rpm and a maximum RCF of 30,000 x g. The LS180 can accommodate various rotor options to accommodate different sample sizes and types.

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LS180 cell line (2 citations)

62 protocols using ls180

Colon Cancer Cell Line Transfection

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Colon cancer cell lines CaR-1, CCK-81, SUN-61, CL-40, LOVO, HT-29, KM-12, CL-11, LS-180, LS-513, and MDST8 were purchased from ATCC company, and all the cell lines above were cultured in Dulbecco's modified Eagle medium (DMEM)+10% fetal bovine serum (FBS) medium. The cultures were maintained at 37°C with 5% CO₂. shRNA and cDNA used in this experiment were synthesized by BGI (Beijing Genomics Institute) company. The transfection of vector was performed using Lipofectamine 2000 reagent (Thermo Fisher Scientific, US); in the transfection, Lipofectamine reagent and vector were first diluted by Opti-MEM medium and then mixed together and cultured for 10 min in room temperature; DNA-lipid complex was then added to cells, and the transfection procedure was finished. And for the cell line experiment, LOVO cells were divided into the LOVO-CON and LOVO MEIS3 shRNA groups, then transfected with CON and MEIS3-shRNA vectors for 48 h, and served for subsequent experiment. In the meantime, SNU-61 cell lines were divided into the CON and MEIS3 groups and transfected with CON and MEIS3 overexpression vectors for 48 h.

Cai P., Xie Y., Dong M, & Zhu Q. (2020). Inhibition of MEIS3 Generates Cetuximab Resistance through c-Met and Akt. BioMed Research International, 2020, 2046248.

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Curcumin Treatment of Cell Lines

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MCF-10A, HL60, SNU-5, LS180, BV2, CHME, and THP1 was purchased from ATCC (American Type Culture Collection) (Manassas, VA) was grown in RPMI media completed with 10% FBS, including penicillin G (70 mg/L), streptomycin (100 mg/L), and NaHCO₃ (3.7 g/L) in an incubator at 37°C, 98% humidity, and 5% CO₂. Treatment of cells with curcumin was dose- and time-dependent and curcumin was dissolved in DMSO (<0.1% DMSO).

Zhang Y., Tu L., Zhou X, & Li B. (2018). Curcumin-Mediated Induction of Apoptosis in Human Glioma CHME Cells. Medical Science Monitor Basic Research, 24, 216-224.

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CTNNB1 Mutational Status in Colorectal Cancer Lines

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Colorectal cancer cell lines DLD1 were purchased from Horizon, HCT116 and LS180 from ATCC. The reported mutational status of CTNNB1 (wt/wt for DLD1, del45S/wt for HCT116, and S45F/S45F for LS180) (https://www.ncbi.nlm.nih.gov/pubmed/24755471) was confirmed by DNA Sanger sequencing. DLD1, HCT116, and LS180 cells were cultured in RPMI medium, DMEM supplemented with 2 mM Ultraglutamine (Lonza), or EMEM, respectively, each containing 10% FCS and 100 U/ml penicillin-streptomycin (Biochrom).

Arnold A., Tronser M., Sers C., Ahadova A., Endris V., Mamlouk S., Horst D., Möbs M., Bischoff P., Kloor M, & Bläker H. (2020). The majority of β-catenin mutations in colorectal cancer is homozygous. BMC Cancer, 20, 1038.

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Colorectal Cancer Cell Viability Assay

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Human colorectal carcinoma LS180, HCT116, LoVo, HT29, HCT15 cell lines and SW620 colon cancer cells derived from metastatic site cell lines were obtained from ATCC (Rockville, MD, USA) and were cultured and grown according to the data sheets.
To evaluate viability, the 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay was used. Briefly, cells were plated at concentrations 2.5 × 10⁴ per well in 24 well plates and treated with OHP and Na⁺ oxalate. At the end of the treatments, the MTT reagent was added to cells at a final concentration of 0.25 mg/mL (Sigma-Aldrich Inc., Milan, Italy), for 90 min at 37 °C. Reactions were stopped and the crystals were solubilized by adding isopropyl alcohol/HCL (1:1; vol:vol, Sigma-Aldrich Inc., Milan, Italy), before reading the absorbance at 570 nm, using the multi-plate reader Victor3 V (PerkinElmer, Milan, Italy). pH determinations with BCECF on LoVo and LS180 was performed as described in the relevant section below.

Riva B., Dionisi M., Potenzieri A., Chiorazzi A., Cordero-Sanchez C., Rigolio R., Carozzi V.A., Lim D., Cavaletti G., Marmiroli P., Distasi C, & Genazzani A.A. (2018). Oxaliplatin induces pH acidification in dorsal root ganglia neurons. Scientific Reports, 8, 15084.

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Colorectal Cancer Cell Lines: Culture Conditions

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The colorectal cancer cell lines HT29 (ATCC^® HTB-38), HT115 (ECACC-cultures 85061104) HCT116 (ATCC^® CCL-247), LS180 (ATCC^® CL-187), Caco-2 (ATCC^® HTB-37), normal colonic epithelium CCD-841-CoN (ATCC^® CRL-1790) and lung fibroblasts MRC5 (ATCC^® CCL-171) were maintained in DMEM media supplemented with 10% fetal bovine serum (Gibco Invitrogen; Paisley, UK). All cultures were maintained at 37°C and at 5% CO2.

Callaghan B., Lydon H., Roelants S.L., Van Bogaert I.N., Marchant R., Banat I.M, & Mitchell C.A. (2016). Lactonic Sophorolipids Increase Tumor Burden in Apcmin+/- Mice. PLoS ONE, 11(6), e0156845.

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Colorectal Cancer Cell Lines and Fibroblasts

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The human CRC cell lines LS180, RKO, SW48, HCT15, HCT116 and Colo320DM were obtained from ATCC (LGC Standards, Wesel, Germany). LS180, RKO and SW48 were kept in MEM (Invitrogen) supplemented with 15% FBS, 1% essential amino acids and antibiotics. HCT15, HCT116 and Colo320DM were kept in RPMI (Invitrogen) supplemented with 10% FBS plus antibiotics. Adult dermal fibroblasts (HDFa, PCS-201-012) and neonatal fibroblasts (HDFn, PCS-201-010) were obtained from ATCC. Fibroblasts were cultured with fibroblast basal medium (FBM, PCS-201-030 from ATCC) supplemented with low serum fibroblast growth kit (ATCC, PCS-201-041). Cells were cultivated at 37 °C in 5% CO₂. Irradiation was done using the RS320 X-Ray machine by XStrahl Ltd. at 300 kV, 10 mA, dose rate 0,9 Gy/min.

Niedermaier B., Sak A., Zernickel E., Xu S., Groneberg M, & Stuschke M. (2019). Targeting ARID1A-mutant colorectal cancer: depletion of ARID1B increases radiosensitivity and modulates DNA damage response. Scientific Reports, 9, 18207.

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Cell Line Authentication and Genetic Manipulation

Cited 1 time

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DLD-1, HT-29, and LS180 cells were purchased from ATCC. LoVo cells were obtained from Dr. Shao-Cong Sun (M.D. Anderson Cancer Center). Cell lines were authenticated utilizing short tandem repeat profiling, routinely tested for mycoplasma, and cultured in RPMI medium supplemented with 10% fetal bovine serum and penicillin/streptomycin at 37°C with 95% humidity and 5% CO₂. Transient transections were performed using Dharmafect Duo (Dharmacon) or jetPRIME (Polypus Transfection). Stable pLKO.1 (control), LGR5, and GPR56 shRNA KD cells were generated by lentiviral infection as previously reported (8 (link),18 (link)). The shRNAs used were, TRCN0000011586 (shLGR5-1), TRCN0000011589 (shLGR5-2), TRCN0000011618 (shGPR56-1), and TRCN0000011619 (shGPR56-2) from GE Dharmacon. Stable DLD-1 cells over-expressing hGPR56 and vector cells were generated as previously described (27 (link)).

Zhang S., Chatterjee T., Godoy C., Wu L., Liu Q.J, & Carmon K.S. (2019). GPR56 drives colorectal tumor growth and promotes drug resistance through upregulation of MDR1 expression via a RhoA-mediated mechanism. Molecular cancer research : MCR, 17(11), 2196-2207.

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Silencing of FOXA1, FORCP, and BRI3BP in Human Cancer Cell Lines

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HEK293T (293T), HCT116, SW48, SW480, RKO, C80, and LS180 cells were purchased from ATCC (Manassas, VA.). SW1222 cells were purchased from Millipore Sigma (St. Louis, MO). All cell lines were maintained in Dulbecco's Modified Eagle's (DMEM) (Thermo Fisher Scientific, Invitrogen) medium containing 10% (v/v) fetal bovine serum (Thermo Fisher Scientific) and 1% penicillin-streptomycin. Cells were cultured at 37°C, 5% CO₂. All cell lines were routinely checked for mycoplasma using the VenorGem Mycoplasma detection kit (Millipore Sigma-Aldrich). OnTarget siRNA SMARTpool for FOXA1, FORCP and BRI3BP were purchased from Thermo Fisher Scientific, Dharmacon. The Allstars Negative control (CTL) siRNAs were purchased from Qiagen. SiRNAs were reverse transfected at a final concentration of 20 nM, using Lipofectamine RNAiMAX (Thermo Fisher Scientific, Invitrogen) as instructed by the manufacturer.

Li X.L., Pongor L., Tang W., Das S., Muys B.R., Jones M.F., Lazar S.B., Dangelmaier E.A., Hartford C.C., Grammatikakis I., Hao Q., Sun Q., Schetter A., Martindale J.L., Tang B., Jenkins L.M., Robles A.I., Walker R.L., Ambs S., Chari R., Shabalina S.A., Gorospe M., Hussain S.P., Harris C.C., Meltzer P.S., Prasanth K.V., Aladjem M.I., Andresson T, & Lal A. (2020). A small protein encoded by a putative lncRNA regulates apoptosis and tumorigenicity in human colorectal cancer cells. eLife, 9, e53734.

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Induction Experiments with LS180 Colon Adenocarcinoma Cells

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The human colon adenocarcinoma cell line LS180 (available at ATCC, Manassas, VA, USA) was used for induction experiments as a surrogate for the intestine being a major site of drug interactions and being an ideal model for investigating pregnane X receptor (PXR) and aryl hydrocarbon receptor (AhR) mediated induction [13 (link),14 (link),15 (link),16 (link),17 (link),18 (link),19 (link),20 (link)]. Cells were cultured under standard cell culture conditions with Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal calf serum, 2 mM glutamine, 100 U/mL penicillin, 100 µg/mL streptomycin sulphate, and 0.1 mM nonessential amino acids.

Weiss J., Theile D., Dvorak Z, & Haefeli W.E. (2014). Interaction Potential of the Multitargeted Receptor Tyrosine Kinase Inhibitor Dovitinib with Drug Transporters and Drug Metabolising Enzymes Assessed in Vitro. Pharmaceutics, 6(4), 632-650.

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Colon Cancer Cell Lines and Genetic Manipulation

Cited 2 times

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LS180, HCT116, and DLD-1 cells were purchased from the ATCC. LoVo cells were obtained from Dr. Shao-Cong Sun (MD Anderson Cancer Center). Cell lines were authenticated utilizing short tandem repeat profiling, routinely tested for mycoplasma, and cultured in RPMI medium supplemented with 10% FBS and penicillin/streptomycin at 37°C with 95% humidity and 5% CO₂. Transient transfections were performed using jetPRIME (Polypus Transfection). LS180 stable LGR5 CRISPR/Cas9 KO clonal lines 1.4 and 1.5 were generated using the lenti-CRISPRv2 vector system as described (12 (link)). LoVo stable shRNA knockdown lines pLKO.1 (shCTL), shLGR5-1, and shLGR5-2 were generated as previously reported (3 (link),13 (link)).

Posey T.A., Jacob J., Parkhurst A., Subramanian S., Francisco L.E., Liang Z, & Carmon K.S. (2023). Loss of LGR5 through therapy-induced downregulation or gene ablation is associated with resistance and enhanced MET-STAT3 signaling in colorectal cancer cells. Molecular cancer therapeutics, 22(5), 667-678.

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