Human colon cancer cell lines Caco-2, SW403, SW1417, COLO 205, HT-29, HCT 116, and RKO were supplied by the American Type Culture Collection (ATCC, Rockville, MD, USA); the HT115 cell line was obtained from Sigma-Aldrich, Milan, Italy. All cell lines were cultured following the manufacturer’s instructions; culture media were supplemented with fetal bovine serum (HyClone, Cramlington, UK), 100 U/mL penicillin, 100 mg/mL streptomycin, and 250 ng/mL Amphotericin-B. The identity of all cell lines was confirmed by short tandem repeat (STR) profiling; cells were routinely screened for mycoplasma contamination with MycoAlert mycoplasma detection kit (Lonza, Milan, Italy).
Ht115
Manufactured by Merck Group
Sourced in United States
The HT115 is a laboratory equipment product from Merck Group. It is designed for general laboratory tasks. The core function of the HT115 is to provide a reliable and consistent heating source for various experimental and analytical procedures. The detailed specifications and intended use of this product are not available for this response.
Automatically generated - may contain errors
Lab products found in correlation
Mycoalert mycoplasma detection kit, by Lonza (1 mentions)
Hct116, by American Type Culture Collection (1 mentions)
Caco 2, by American Type Culture Collection (1 mentions)
Colo205, by American Type Culture Collection (1 mentions)
Sw1417, by American Type Culture Collection (1 mentions)
Sw403, by American Type Culture Collection (1 mentions)
Colo201, by American Type Culture Collection (1 mentions)
Alamarblue buf012b, by Bio-Rad (1 mentions)
Ht 29, by CLS Cell Lines Service (1 mentions)
Sw620, by CLS Cell Lines Service (1 mentions)
Ht 29, by Merck Group (1 mentions)
Sb431542, by Merck Group (1 mentions)
Sw620, by Merck Group (1 mentions)
Pen strep, by Thermo Fisher Scientific (1 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
Phenol red, by Merck Group (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Snu c4, by Korean Cell Line Bank (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
4 protocols using ht115
1
Culturing Human Colon Cancer Cell Lines
2
PEDF Modulation of Colorectal Cancer Cells
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Human colorectal cancer cell lines RKO, COLO-201, LSI74T and colorectal fibroblast cell line CCD-33C0 were purchased from American Type Culture Collection (ATCC) (Rockville, Maryland, USA). Human colorectal cancer cell lines HT115, HRT-18 were purchased from Sigma-Aldrich (Poole, Dorset, UK). The cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM)-F12 medium supplemented with 10% foetal calf serum and antibiotics. Polyclonal rabbit anti-PEDF was obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, Texas, USA). Recombinant PEDF (rhPEDF) was purchased from R&D Systems Europe Ltd (Abingdon, Oxfordshire, UK) and cellular functional assays were performed at doses of 10 ng/ml, 50 ng/ml and 100 ng/ml versus control medium. HT115 and HRT-18 were used as representative colorectal cancer cell lines for cellular functional assays. Unless otherwise stated, other materials and reagents were purchased from Sigma-Aldrich Ltd (Poole, Dorset, UK).
3
Colorectal Cancer Cell Lines Inhibition
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Human colorectal cell lines HT-29, HT115, and SW620 were purchased from CLS Cell Lines Service GmbH (Eppelheim, Germany). The cell line was incubated in Dulbecco's modified Eagle's medium supplemented with d -glucose 4500 mg/l, 4 mM l -glutamine and 110 mg/l sodium pyruvate, 10% fetal bovine serum, 1x penicillin–streptomycin (Pen-Strep) and non-essential amino acids (all purchased from Gibco-Invitrogen, Waltham, MA, USA). Colon cancer cell line HT-29, HT115, and SW620 were treated with 5 μM DZNep small-molecule inhibitor of EZH2 (Sigma, St. Louis, MO, USA). Pharmacological inhibition of Wnt pathway was conducted using XAV939 (0.25 and 1 μM ) and IWP-2 (1 and 5 μM ). Inhibition of TGF-β pathway was conducted as we previously described using 10 μM SB-431542 (Sigma, St. Louis, MO, USA).34 (link) Assays were carried out with appropriate controls such as dimethyl sulfoxide. Briefly, 10 000 cells were cultured in a 96-well plate and cell viability was measured at the indicated time points using the alamarBlue (BUF012B; AbD Serotec, Kidlington, UK) assay.
4
Characterization of Colorectal Cancer Cell Lines
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All cells were grown at 37 C and 5% CO 2 . Tumor cell lines NCI-H508, LS-180, and SW48 were obtained from the American Type Culture Collection (ATCC), SNU-C4 from the Korean Cell Line Bank (Seoul National University), and HT115 from Sigma-Aldrich. Cell lines were analyzed for authenticity by the respective cell bank or upon receipt and tested negative for Mycoplasma contamination. In the absence of phenotypic or growth changes, cells were not further authenticated or tested for Mycoplasma. All cell lines were used within 2 months of thawing. Cells were cultured in RPMI-1640 (Gibco) with 10% fetal bovine serum (FBS), GlutaMAX (Gibco), penicillin/ streptomycin (Corning), nonessential amino acids (Thermo Fisher Scientific), and sodium pyruvate (Thermo Fisher Scientific) or DMEM (Biochrom) þ 15% FBS. Chinese hamster ovary (CHO) cells overexpressing membrane-bound mucins were cultured in HyClone PF CHO Media (GE) containing phenol red (Sigma-Aldrich) and penicillin/streptomycin (Corning). Generation of cells with CRISPR knockout of MUC12 and transfected CHO cells is described in Supplementary Methods.
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