Isolation and culture of bone marrow-derived BCR/ABL+ MSCs from CML patients were performed as described previously with some modifications19 (link)20 (link)21 (link). Briefly, mononuclear cells were separated by a Ficoll-Paque gradient centrifugation (specific gravity 1.077 g/mL; Nycomed Pharma AS, Oslo, Norway) and the sorted cells were plated at concentration of 1 cell/well by limiting dilution in a total of 96 × 10 wells coated with fibronectin (Sigma, St Louis, MO) and collagen (Sigma) for each patient. Culture medium was Dulbecco modified Eagle medium and Ham F12 medium (DF12) containing 40% MCDB-201 medium complete with trace elements (MCDB) (Sigma), 2% fetal calf serum (FCS; Gibco Life Technologies, Paisley, United Kingdom), 1 × insulin transferrin selenium (Gibco Life Technologies), 10−9 M dexamethasone (Sigma), 10−4 M ascorbic acid 2-phosphate (Sigma), 20 ng/mL interleukin-6 (Sigma), 10 ng/mL epidermal growth factor (Sigma), 10 ng/mL platelet-derived growth factor BB (Sigma), 50 ng/mL fetal liver tyrosine kinase 3 (Flt-3) ligand (Sigma), 30 ng/mL bone morphogenetic protein-4 (Sigma), 100 U/mL penicillin and 100 ug/mL streptomycin (Gibco Life Technologies) at 37 °C and a 5% CO2 humidified atmosphere. Culture media were changed every 4 to 6 days.
Ham f12 medium
Manufactured by Merck Group
Sourced in United States
Ham F12 medium is a cell culture medium used for the growth and maintenance of various cell types, including mammalian cells. It provides a balanced mixture of nutrients, vitamins, and other essential components required for cell proliferation and survival. The core function of Ham F12 medium is to support the in vitro cultivation of diverse cell lines.
Automatically generated - may contain errors
Lab products found in correlation
Fetal calf serum (fcs), by Thermo Fisher Scientific (3 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Horse serum, by Thermo Fisher Scientific (2 mentions)
Fibronectin, by Merck Group (1 mentions)
Insulin transferrin selenium, by Thermo Fisher Scientific (1 mentions)
Mcdb 201 medium, by Merck Group (1 mentions)
Interleukin 6 (il 6), by Merck Group (1 mentions)
Platelet derived growth factor bb, by Merck Group (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Ascorbic acid 2 phosphate, by Merck Group (1 mentions)
Epidermal growth factor (egf), by Merck Group (1 mentions)
D glucose, by Merck Group (1 mentions)
Sodium pyruvate, by Merck Group (1 mentions)
Sodium bicarbonate, by Merck Group (1 mentions)
Foetal calf serum, by Merck Group (1 mentions)
Williams e medium, by Merck Group (1 mentions)
Caco 2, by RIKEN Cell Bank (1 mentions)
10 protocols using ham f12 medium
1
Isolation and Culture of BCR/ABL+ MSCs from CML Patients
2
BHK-21 Cell Culture Protocol
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BHK-21 cells provided by Pasteur Institute, (FR) were cultured in Dulbecco’s Modified Eagle’s medium (D-MEM, Sigma-Aldrich, US) and HAM F-12 medium (Sigma-Aldrich, US) supplemented with 0.055% sodium pyruvate (Sigma-Aldrich, US), 2.44% sodium bicarbonate (Merck, US), 1.5% D-(+)-glucose (Sigma-Aldrich, US), 0.05% gentamicin sulfate (Inlab, BR) and 5% fetal bovine serum (FBS) (Laborclin, BR). Cells were transferred twice per week in T75 flasks at a density of 1.5 x 105 cells/cm2 and incubated in a humidified incubator at 37°C with 5.0% CO2, unless stated otherwise.
3
Detailed Cell Culture Media Reagents
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ABZ and RCB was purchased from Sigma-Aldrich (St. Louis, MO, USA). FLU was obtained from Janssen Pharmaceutica (New Brunswick, NJ, USA). Liquid sterile-filtered medium RPMI-1640 medium, HAM F12 medium, Williams’ E medium, foetal calf serum and other chemicals (UHPLC, MS or analytical grade) were obtained from Sigma-Aldrich (St. Louis, MO, USA).
4
Culturing Colon Cancer SW-480 Cells
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Epithelial colon cancer cells, SW-480 cell line (CLS GmbH, Kirkel, Germany) passage No 49–51, were used in the experiments. The cells were cultured in Ham F-12 medium (Sigma Aldrich, Baden-Württemberg, Germany) supplemented with 10% fetal calf serum (Gibco, North Lincolnshire, UK), streptomycin, amphotericin B and penicillin (Anti-Anti, Gibco, North Lincolnshire, UK) and maintained at 37 °C in an atmosphere of 10% CO2 and 95% humidity in a CO2 incubator (MRC, Holon, Israel). The cell culture passage was performed upon reaching 80% confluence.
5
Cell Culture Conditions for Drug-Resistant Cells
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DLD-1 cells were purchased from the Health Science Research Resources Bank (Osaka, Japan). LoVo, Colo-205 and Caco-2 cells were purchased from Riken Cell Bank (Ibaraki, Japan). HT-29 cells were purchased from DS Pharma Biomedical (Osaka, Japan). These cells were cultured in RPMI1640 medium (Sigma, St Louis, MO, USA), HamF12 medium (Sigma) or McCoy’s 5A medium (Sigma) supplemented with 10% fetal bovine serum (Gibco, Carlsbad, CA, USA), 100 μg/mL penicillin (Gibco), 100 U/mL streptomycin (Gibco) and 25 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (pH 7.4; FUJIFLIM Wako, Tokyo, Japan) in an atmosphere containing 5% CO2.
LoVo and Colo-205 cells with acquired resistance to PD0325901 (LC Laboratories, Woburn, MA, USA) or trametinib (LC Laboratories) (LoVo/PR, Colo-205/PR and LoVo/TR) were generated as previously described [37 (link),51 (link),52 (link),53 (link),54 (link),55 (link)].
LoVo and Colo-205 cells with acquired resistance to PD0325901 (LC Laboratories, Woburn, MA, USA) or trametinib (LC Laboratories) (LoVo/PR, Colo-205/PR and LoVo/TR) were generated as previously described [37 (link),51 (link),52 (link),53 (link),54 (link),55 (link)].
6
Fetal Bovine and Horse Serum Assay
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Fetal bovine serum and horse serum were obtained from Gibco (UK). HAM-F12 medium,
phosphate buffered saline (PBS), streptomycin, penicillin, gentamicin, methylmercury
chloride (MeHgCl, 99.8%), N-nitro-L-arginine (L-NARG),
3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT), and all reagents
for radioimmunoassays were purchased from Sigma-Aldrich (USA).
phosphate buffered saline (PBS), streptomycin, penicillin, gentamicin, methylmercury
chloride (MeHgCl, 99.8%), N-nitro-L-arginine (L-NARG),
3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT), and all reagents
for radioimmunoassays were purchased from Sigma-Aldrich (USA).
7
Vacuolation Quantification in RK13 Cells
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RK13 cells (American Type Culture Collection CCL-37) were maintained at 37°C and 5% carbon dioxide in Ham F12 medium supplemented with 2 mmol/L L-glutamine and 10% fetal bovine serum (Sigma-Aldrich). Bacterial water extracts were prepared as described elsewhere [10 (link)] and applied at 75 µg/mL to 1 × 104 cells per well in 96-well plates in the presence of 10 mmol/L ammonium chloride. After 4 hours, the extent of vacuolation was quantified with light microscopy and expressed as the mean proportion of vacuolated cells per field (≥100 cells assessed per experimental condition, 4 fields per condition, and duplicate independent experiments).
8
Culture and Maintenance of Bronchial Epithelial Cell Lines
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The normal human bronchial epithelial cell line BEAS-2B (50 (link)) was purchased from the American Type Culture Collection. The original BEAS-2B line and the reporter line BEAS-2B.R05Z were routinely cultured in a 1:1 Dulbecco's modified Eagle's medium (DMEM)–Ham F-12 medium (Sigma) mixture supplemented with 2% newborn calf serum (NCS; HyClone); 2 mg/liter insulin, 2 mg/liter transferrin, and 2 μg/liter selenite (2-ITS) (all from Sigma); and a 1% (vol/vol) penicillin-streptomycin cocktail (HyClone). The immortalized human cystic fibrosis bronchial epithelial line CFT-1 and a matching normal cell line, HBE-1 (51 (link)), were obtained from Kerafast. CFT-1 and HBE-1 cells were cultured in bronchial epithelial cell growth medium (Lonza). The cells were subcultured at a 1:5 ratio upon reaching near confluence. The standard culturing conditions for all cells were 37°C with 5% CO2 and 100% humidity.
9
Lymphocyte Oxidative Stress Analysis
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Blood samples were obtained from healthy female donors aged 20-40 years. They were collected by venous puncture with pre-heparinized syringes and cultured in Falcon T25 flasks (Nunc, Denmark) containing HAM F12 medium (Sigma-Aldrich, St. Louis, MO, USA) with antibiotics (60 UI penicillin, 50 μg/ml streptomycin) (Bagó, Buenos Aires, Argentina) and without fetal bovine serum. Lymphocytes were stimulated with 100 μg/ml phytohemagglutinin (Gibco Thermo Fisher Scientific, Buenos Aires, Argentina). All cultures were performed in a pool of gene samples at 37 °C in 5% CO 2 for 7 days. Cultures were performed for 7 days without changing the medium to evaluate the chronic and cumulative effect of daily ferrous sulfate supplementation. Changing the medium would mean to replenish the ferrous sulfate supplemented daily, causing a new metabolic peak which may skew results. Oxidative stress was assessed in samples cultured in HAM F12 medium without phenol red (EMEVE media) (Lobov Buenos Aires, Argentina).
10
Long-term Estrogen Exposure in GH3B6 Cells
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The rat somatolactotroph pituitary adenoma GH3B6 cell line was cultured in HAM-F12 medium (Sigma-Aldrich) supplemented with 5% v/v FCS and 12% v/v horse serum (Invitrogen), as previously described (Petiti et al. 2010) (link). After 3 days of culture and 70% of confluence in order to recreate estrogenic doses administration at E40-E60, cells were exposed to 17b-estradiol (E 2 ; 4 nM; Sigma-Aldrich) or vehicle for 120 h and then submitted to the cytochemical detection of SA-b-gal staining.
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