C6 rat glioma cells (ATCC #CCL-107), MDA-MB-231 cells (ATCC #HTB-26), and HEK293 cells (ATCC #CRL-1573) were maintained as a monolayer culture on tissue culture dishes at 37 °C, 5% CO2, 100% humidity in Dulbecco’s modified Eagle’s medium supplemented with 10% heat-inactivated fetal bovine serum and antibiotics. S1P and CYM-5478 were solubilized with bovine serum albumin (0.1% final concentration) prior to treatment.
Ccl 107
Manufactured by American Type Culture Collection
Sourced in United States
CCL-107 is a human breast cancer cell line derived from a pleural effusion. It is maintained in culture and can be used for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Male wistar rats, by Charles River Laboratories (2 mentions)
Dbcamp, by Merck Group (1 mentions)
Crl 1690, by American Type Culture Collection (1 mentions)
Tc plate 24 well, by Greiner (1 mentions)
Nucleofector 2b device, by Lonza (1 mentions)
Bestatin, by Merck Group (1 mentions)
Htb 22, by American Type Culture Collection (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Trypan blue, by Thermo Fisher Scientific (1 mentions)
Fungizone, by Thermo Fisher Scientific (1 mentions)
Tryple express enzyme solution, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Rat c6 glioma cells, by American Type Culture Collection (1 mentions)
Ccl 34, by American Type Culture Collection (1 mentions)
Crl 1439, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Lonsera (1 mentions)
Htb 14, by American Type Culture Collection (1 mentions)
15 protocols using ccl 107
1
Culturing Glioma, Breast, and Kidney Cells
2
Cell Culture Biocompatibility on Membranes
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Membrane samples of 14 mm diameter were glued to glass coverslips to prevent floating, sterilized with 70% v/v ethanol (EtOH, > 99.8%, Honeywell) solutions and UV light, and washed with sterile PBS to remove EtOH traces, all in a laminar flow cabinet.
Cell biocompatibility and astrocytic differentiation on PCL and PCL/GBN membranes were carried out with C6 rat glioma cells (CCL-107™, ATCC®). C6 cells were cultured in DMEM, enriched with 10% FBS, 1% of non-essential amino acids (NEAA) and 1% of penicillin/streptomycin (P/S) at 37ºC in a humid atmosphere containing 5% CO2. All supplements were acquired from Gibco™. Culture medium was changed every 2 days until 80% of cell confluence. Then, C6 cells were uniformly seeded (3.2 × 104 cells per cm2) on flat membranes. After 24 h, astrocytic differentiation was induced for 5 days using 0.5% FBS in DMEM containing 1 mM of dibutyryl-cAMP (dbcAMP, Sigma Aldrich). Results were compared with glass coverslips as positive control.
Cell biocompatibility and astrocytic differentiation on PCL and PCL/GBN membranes were carried out with C6 rat glioma cells (CCL-107™, ATCC®). C6 cells were cultured in DMEM, enriched with 10% FBS, 1% of non-essential amino acids (NEAA) and 1% of penicillin/streptomycin (P/S) at 37ºC in a humid atmosphere containing 5% CO2. All supplements were acquired from Gibco™. Culture medium was changed every 2 days until 80% of cell confluence. Then, C6 cells were uniformly seeded (3.2 × 104 cells per cm2) on flat membranes. After 24 h, astrocytic differentiation was induced for 5 days using 0.5% FBS in DMEM containing 1 mM of dibutyryl-cAMP (dbcAMP, Sigma Aldrich). Results were compared with glass coverslips as positive control.
3
Evaluation of Nanoparticle Cytotoxicity
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The influence of the Au-CuO and CuO-ZnO nanoparticles compounds on the viability of cells in vitro was evaluated with use two established cell lines received from American Type Culture Collection (Manassas, VA, USA). The rat brain glioma cell line C6 (ATCC® CCL-107™) was cultured in DMEM medium containing 10% fetal bovine serum, 50 U-mL penicillin, 100 μg-mL streptomycin, and 25 μg-mL amphotericin B. The human glioblastoma cell line T98G (ATCC® CRL-1690™) was grown in EMEM medium supplemented as mentioned above. Cells were kept at 37 °C in a 5% CO2 atmosphere in the incubator with increased humidity up to 85% confluency. Cells were plated in quantity 5 × 105 cells per well and cultured in the plastic 24-Well flat-bottom plates (TC-PLATE 24 well, Greiner).
4
Nucleofection of C6 Glioma Cells
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C6 cells were obtained from American Type Culture Collection (CCL-107, ATCC, RRID:CVCL_0194 ) and cultured in F-12k-based medium (2.5% bovine serum, 12% horse serum). At each passage, cells were trypsinized for 1–3 min (0.25% trypsin and 1 mM EDTA in PBS pH 7.4) at room temperature. After each passage remaining cells were processed for nucleofection (2 × 106/group). Cell pellets were resuspended in nucleofection buffer (5 mM KCl, 15 mM MgCl, 15 mM HEPES, 125 mM Na2HPO4/NaH2PO4, 25 mM Mannitol) and electroporated with 3.4 μg plasmid DNA per group. Nucleofector™2b device (Lonza) was used according to the manufacturer's instruction (C6, high efficiency protocol). Nucleofection groups were diluted with 500 μl media respectively and plated in triplicates in 24-well plates (∼ 666 667 cells/well). Plates underwent a full media change 4–6 h after nucleofection and were imaged and frozen for downstream processing after 16 h.
5
Investigating Acanthamoeba Virulence Mechanisms
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To find out the probable mechanisms of virulence in vitro, the experiments following the previous study was conducted using a rat glial C6 cell line (ATCC® CCL-107™) to incubate Acanthamoeba and secreted proteins [18 (link)]. Rat glial C6 cells were cultured in 10 mL Dulbecco’s minimal essential medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and antibiotics (50 μg/mL streptomycin and 100 U/mL of penicillin) at 37 °C incubate with 5% CO2. Rat glial C6 cells were cultured 3 × 105 in 24-well plate cell culture dishes. After 24 h of growth, Acanthamoeba live cells at a concentration of 3 × 105 were co-cultured with C6 cells. Acanthamoeba, at a density of 1.0 × 106 parasites/mL in PBS for 24 h, as well as their secreted proteins were collected. Acanthamoeba secreted proteins (150 μg) were co-incubated with rat glial C6 cell for 2 h followed by time-lapse microscopy observations and CPE assays. The effects of 100 μM aminopeptidase inhibitor (bestatin; Sigma-Aldrich, Darmstadt, Germany) and 15 μg antibody of M20/M25/M40 on C6 cell co-incubation with Asp for 2 h were monitored by CPE assay and time-lapse observation.
6
Culturing C6 Glioma and MCF-7 Cells
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7
C6 Glioma Cell Culture Protocol
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The C6 glioma cell line (CCL-107; American Type Culture Collection, Manassas, VA, USA) was used in the present study. In comparison with other experimental glioma tumors (CNS1, RT-2, BT4C, F98, RG2, T9), С6 has the largest proportion of CSCs (12 (link)). The cells were cultivated at 37°C with 5% CO2 in 25-cm2 vials filled with Dulbecco's modified Eagle medium (DMEM) medium containing 10% fetal bovine serum (FBS) and Antibiotic-Antimycotic (100X), containing 10,000 IU/ml penicillin, 10,000 µg/ml streptomycin and 25 µg/ml fungizone (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA). The culture medium was renewed every 3 days. After reaching 95% confluence, the cells were treated with TrypLE Express Enzyme solution (Gibco; Thermo Fisher Scientific, Inc.; 12604-021) for 7 min at 37°C, placed into a 15-ml vial and centrifuged for 3 min at 120 × g. The supernatant was removed, DMEM + 10% FBS + 100X Antibiotic-Antimiotic fresh medium was added to the pellet and the culture was transferred into a new 25-cm2 culture vial.
Following suspension in fresh medium, the cells were counted using a hemocytometer and their viability was determined by staining with 0.4% trypan blue (Gibco; Thermo Fisher Scientific, Inc.; 15250-061). Prior to implantation into rat brains, C6 cells were assigned immunocytochemical characteristics, as follows.
Following suspension in fresh medium, the cells were counted using a hemocytometer and their viability was determined by staining with 0.4% trypan blue (Gibco; Thermo Fisher Scientific, Inc.; 15250-061). Prior to implantation into rat brains, C6 cells were assigned immunocytochemical characteristics, as follows.
8
Culturing Rat Alveolar Macrophages and Glioma Cells
9
Culturing Rat Glioma C6 Cells
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The rat glioma C6 cell line was obtained from the American Type Culture Collection (ATTC® CCL-107™, Manassas, VA). Cells were cultured in Dulbecco´s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum, 100 U/mL penicillin and 100 g/mL streptomycin. Cells were grown in a humidified atmosphere with 5% CO2 at 37°C. When the culture reached a confluence greater than 90%, it was separated from the substrate to perform cell count and viability tests with trypan blue. The minimum viability of the suspension (1x105 cell/μL) of successful cultures was set to 90%.
10
Orthotopic C6 Glioma Model in Rats
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Male Wistar rats weighing 300–400 g (N = 33) were used in this study (Charles River Canada, age 8 to 10 weeks at surgery). The animals were anaesthetized with 2% isoflurane during all procedures. C6 glioma cells (CCL-107, American Type Culture Collection, Manassas, VA) were cultured in F12k 15% horse serum, 2.5% bovine serum, and 1% penicillin-streptomycin. For the implantation of C6 glioma cells, each animal was secured into a stereotactic surgical frame. After scalp incision at midline and exposing the bregma, a 1 mm diameter burr hole was drilled at 1 mm anterior and 3 mm right of the bregma. A total of 106 C6 glioma cells were slowly injected for 5 minutes at a depth of 3–4 mm from the skull surface, which corresponded to the location of the caudate putamen [20] . The burr hole was sealed with bone wax, and the scalp was closed with sutures.
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