The human colorectal cancer cell lines LoVo and HCT116 were purchased from the American Type Culture Collection (USA) and cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum, 100 U/mL penicillin, and 100 g/mL streptomycin. When the confluence of LoVo and HCT116 cells reached 90%, we treated LoVo with 300 μM of CoCl2 (Sigma-Aldrich, St. Louis, MO, USA) for 72 h and HCT116 with 450 μM of CoCl2 for 48 h. Most diploid LoVo and HCT116 cells died following CoCl2 treatment, whereas scattered PGCCs survived. A PGCC was defined as a tumor cell with a nucleus at least 3 times larger than that of a diploid tumor cell 6 (link), 12 (link), 14 (link).
Cocl2
Manufactured by Merck Group
Sourced in United States, Germany, China, United Kingdom, Israel, Switzerland, Canada
CoCl2 is a chemical compound used in various laboratory applications. It serves as a desiccant and can be used as an indicator for detecting moisture levels. The core function of CoCl2 is to provide a visual indication of the presence or absence of water.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (52 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (18 mentions)
Zncl2, by Merck Group (17 mentions)
Nicl2, by Merck Group (16 mentions)
Mncl2, by Merck Group (15 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (15 mentions)
Fecl3, by Merck Group (14 mentions)
Penicillin, by Thermo Fisher Scientific (14 mentions)
Mgcl2, by Merck Group (13 mentions)
Streptomycin, by Thermo Fisher Scientific (13 mentions)
Cucl2, by Merck Group (12 mentions)
Fecl2, by Merck Group (11 mentions)
Mg132, by Merck Group (11 mentions)
Sodium chloride (nacl), by Merck Group (10 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (10 mentions)
Potassium chloride (kcl), by Merck Group (9 mentions)
Cdcl2, by Merck Group (8 mentions)
Cacl2, by Merck Group (8 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (8 mentions)
Sodium hydroxide (naoh), by Merck Group (7 mentions)
364 protocols using cocl2
1
Generating Polyploid Giant Cancer Cells
2
Hypoxia-Induced Breast Cancer Stem Cell Study
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CoCl2 (Sigma-Aldrich; Merck KGaA) was dissolved in DMEM at a concentration of 100 mM. To mimic hypoxic conditions, 100 mM CoCl2 was added to the three breast cancer cell lines for 24, 48, and 72 h, and BCSCs were transfected with miR-137 mimics for 12 and 24 h for further examination. LW-6 (cat. no. S8441; Selleck Chemicals) was dissolved in dimethyl sulfoxide (DMSO) at a concentration of 25 mM. BCSCs were treated with DMEM containing 10 ng/ml LW-6 or DMSO (control group) for 12 or 24 h to explore the role of miR-137 mimics.
3
Evaluating Glioma Cell Viability
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The U251 cell glioma line was obtained from National Collection of Authenticated Cell Cultures,RRID:CVCL_0021, China, 2018. Cells were divided into three groups: blank control group, MgCl2 group, and CoCl2 group. Briefly, 2000 cells per well were inoculated into 96-well plates, with three replicates in each group. The blank control group was cultured in a normal culture medium without additional treatment; the MgCl2 group was cultured in a normal culture medium containing 400 μmol/L MgCl2, and the CoCl2 group in 400 μmol/L CoCl2 (Sigma, USA) culture medium. After 24 hours, 20 μl of CCK-8 solution (Yeasen Biotechnology, Shanghai, China) was added to each well, and the culture plate was incubated in the incubator for an additional 2 hours. The light absorption value of each well was measured on the microplate reader (OD450 nm), the results were recorded, and the cell viability value was calculated. Taking the CoCl2 group as an example, the cell viability value% = [OD (CoCl2 group) - OD (blank)] / [OD (blank control group) - OD (blank)] × 100%.
4
Detecting MPTP Opening Dynamics
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Fluorescent Calcein AM (Life Technology, New York, America; Cat.C3099 Lot: 1,311,548) and CoCl2 (Sigma, Lot: 232,696 No: BCBG0246V) markers were employed to detect the dynamic changes of MPTP opening using FC. At 4, 24, 48, 72, 96, and 120 h after infection, chick embryo caecal epithelial cells were harvested using 0.25% trypsin, rinsed in PBS, centrifuged at 600 g for 5 min, suspended in 200 μl of binding buffer (Sigma, California, America), and incubated with Calcein AM and CoCl2 (15 min, 37 °C, in the dark). Subsequently, 250 μl of binding buffer was added to the cells, and the mixture was subjected to FC analysis. Flow cytometry (American BD, FACSCalibur) was performed as previously described [21 ]. The results were analysed using CellQuest software. Fluorescent intensity reflects a change in MPTP opening [22 (link)].
5
Hypoxia and FAK Inhibition Impact on BMSCs
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1×105 BMSCs or 7×105 M210B4 cells/well were cultured overnight in 12-well plates (BD Falcon, Bedford, MA) in their respective media. The cells were treated with fresh medium supplemented with 100 μM of cobalt chloride (CoCl2) (Sigma-Aldrich, St. Louis, MO) for 48 h, following which the medium was replaced with fresh medium without CoCl2 (CoCl2-BMSCs) and the cultures were incubated under normoxia (21% oxygen, which is the standard cell culture condition). In hypoxia treatments, the cells were either continuously cultured under 1% oxygen (hypoxic-BMSCs) or under normoxia after preincubation for 48 h under hypoxia (BMSCs-48 h-hypoxia). For the treatment of focal adhesion kinase (FAK) inhibitor PF573228 (PF228; Sigma-Aldrich), overnight-cultured BMSCs (1×105/well of 12-well plate) were pretreated with 300 nM PF228 for 2 h, followed by the addition of 100 μM of CoCl2 for 48 h. Both compounds were removed before seeding the HSCs.
The cell viability in all cases was >98% as determined by Trypan Blue dye exclusion method.
The cell viability in all cases was >98% as determined by Trypan Blue dye exclusion method.
6
Neuroprotective Effects of CLO and STO
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HT22 cells were randomly divided into 10 groups (n = 6 per group): a CoCl2 group (CoCl2) as the model group, a normal control group (Con), four CLO and four STO groups (100, 200, 400, and 600 μg/mL). STO was administered 4 hours after reperfusion. The dose chosen in this study was based on our preliminary studies. In the CLO and STO groups, HT22 cells were incubated with different concentrations of CLO and STO. In vitro ischemia and reperfusion treatment were performed as described previously (Yang et al., 2015). HT22 cells in the CoCl2 group were incubated with 500 μM CoCl2 (Sigma) for 16 hours and culture medium was removed thereafter. Meanwhile, DMEM with 10% FBS were added. The control group was treated with DMEM with 10% FBS and was maintained under the same conditions.
7
CoCl2 Cytotoxicity Evaluation in LoVo Cells
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The effects of CoCl2 on the viability of LoVo cells were detected using an MTT assay. The LoVo cells, at a density of 5×103cells/well, were cultured for 24 h and then treated with 100, 200, 300, 400 or 500 µmol/l CoCl2 (Sigma-Aldrich) for 1, 3, 5 or 7 days. The experiment was performed in triplicate. Subsequently, the plates were washed extensively with serum-free DMEM to remove CoCl2 and dead cells, and were exposed to 20 µl (5 g/l) MTT (Sigma-Aldrich) for 4 h. The resulting formazan crystals were dissolved in 200 µl dimethyl sulfoxide (Sigma-Aldrich) and the absorbance was measured at 490 nm with a microplate reader (Victor3; PerkinElmer, Inc., Waltham, MA, USA). The cytotoxicity of CoCl2 to LoVo cells was evaluated by determination of the survival rate of LoVo cells (calculated as A490 of treatment group/A490 of untreated group).
8
Hypoxia and CoCl2 Regulate Esophageal Cancer
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Eca109 and KYSE450 esophageal cancer cell lines were cultured at 2×10
6 cells/mL in six-well plates in DMEM with 10% FBS and antibiotics (10,000 U/mL penicillin and 10 μg/mL streptomycin) at 37°C in a humidified atmosphere containing 5% CO
2 (normoxia) or in a hypoxial tank containing 94% N
2, 5% CO
2 and 1% O
2 (hypoxia) for 4 h. Then cells were harvested for RT-PCR and western blot analysis. For CoCl
2 treatment, Eca109 and KYSE450 cells were cultured in DMEM with or without 200 μM CoCl
2 (Sigma-Aldrich) for 48 h prior to RT-PCR and western blot analysis.
6 cells/mL in six-well plates in DMEM with 10% FBS and antibiotics (10,000 U/mL penicillin and 10 μg/mL streptomycin) at 37°C in a humidified atmosphere containing 5% CO
2 (normoxia) or in a hypoxial tank containing 94% N
2, 5% CO
2 and 1% O
2 (hypoxia) for 4 h. Then cells were harvested for RT-PCR and western blot analysis. For CoCl
2 treatment, Eca109 and KYSE450 cells were cultured in DMEM with or without 200 μM CoCl
2 (Sigma-Aldrich) for 48 h prior to RT-PCR and western blot analysis.
9
Hypoxia-Induced Cell Viability Assay
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The cells were seeded in 96-well culture plates at a density of 1 × 104 cells/well for 24 h. The culture medium was changed to fresh MEM containing 2% FBS and 1% antibiotics, and for hypoxic preconditioning COCl2 was added to the complete medium at a concentration of 100 μM; COCl2 was purchased from Sigma (Gabriella et al. 2018 ).
After 24 h, the medium was changed with a new one supplemented with 0.5 mg/ml of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) for 3 h at 37 °C. The formazan produced was dissolved by solvent solution (0.1 N HCl in isopropanol), and the optical density was read at 570 nm by a microplate reader (Model 680, Bio-Rad Lab Inc., CA, USA) (Lan et al. 2015 (link)).
After 24 h, the medium was changed with a new one supplemented with 0.5 mg/ml of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) for 3 h at 37 °C. The formazan produced was dissolved by solvent solution (0.1 N HCl in isopropanol), and the optical density was read at 570 nm by a microplate reader (Model 680, Bio-Rad Lab Inc., CA, USA) (Lan et al. 2015 (link)).
10
Propofol Modulates CoCl2-Induced BV2 Cells
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To determine the appropriate treatment condition, BV2 cells were treated with 500 μM CoCl2 (sigma) for 0, 1, 2, 4, 8 and 12 h respectively. Cells culture in DMEM, without any treatment, served as the control group. The time with maximal effects on TNF-α production was used as the appropriate treatment condition. Then cells were pre-treated with different concentrations of propofol (sigma) (5 μmol/L, 25 μmol/L, 50 μmol/L and 100 μmol/L), followed by CoCl2 (500 μM, 8 h) treatment, and the concentration with maximal protective effects was determined. In the following experiments, the optimal concentration of CoCl2 and propofol were used to investigate the potential mechanisms.
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