Trypsin versene mixture
Manufactured by Lonza
Sourced in United States
Trypsin-versene mixture is a laboratory reagent used for the dissociation and detachment of adherent cells from cell culture surfaces. It contains the enzymes trypsin and EDTA (versene) which work together to break down the extracellular matrix and cell-cell adhesions, allowing the cells to be harvested and subcultured.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Penicillin and streptomycin p s, by Lonza (2 mentions)
Heparg cells, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
L glutamic acid, by Merck Group (1 mentions)
T 75 tissue culture flasks, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
Taxol, by Merck Group (1 mentions)
Rpmi 1640 cell culture medium, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Lonza (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Trifluoroacetic acid, by Merck Group (1 mentions)
Trans retinoic acid, by Merck Group (1 mentions)
Cis platinum, by Merck Group (1 mentions)
Trioctylphosphine oxide, by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Thermo Fisher Scientific (1 mentions)
Sodium chloride (nacl), by Thermo Fisher Scientific (1 mentions)
11 protocols using trypsin versene mixture
1
Cell Culture Reagents and p53 Inhibitor
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l -Glutamic acid, estrogens and their metabolic derivatives, and fetal bovine serum (FBS) are obtained from Sigma (St. Louis, MO). Dullbecco’s modified Eagle’s medium (DMEM) is obtained from Life Technology (Rockville, MD). The antibiotics solution (containing 10,000 U/mL penicillin and 10 mg/mL streptomycin) is obtained from Invitrogen (Carlsbad, CA), and trypsin-versene mixture (containing 0.25% trypsin and 0.02% EDTA) from Lonza Walkersville (Walkersville, MD). Pifithrin-α (PFT-α, p53 inhibitor) is obtained from Calbiochem (San Diego, CA).
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2
Culturing HEK293, Huh7, and LH86 Cells
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Human embryonic kidney (HEK293) and HCC (Huh7 and LH86) cells were described previously.7 (link),37 (link) All cells were maintained in complete Dulbecco’s Modified Eagle Medium (Mediatech, Manassas, VA) supplemented with 10% heat-inactivated fetal bovine serum (Sigma-Aldrich, St. Louis, MO), 1% penicillin and streptomycin (P/S, Lonza, Walkersville, MD). Cells were grown as adherent culture in a humidified atmosphere at 37 °C in 5% CO2 and were subcultured after treatment with trypsin–versene mixture (Lonza, Walkersville, MD) for 2–5 minutes at room temperature (RT), washed and resuspended in complete Dulbecco’s Modified Eagle Medium. Recombinant AAV2 and pHelper plasmids have been described previously.38 (link) A recombinant plasmid containing the WT AAV3 genome was a kind gift from Dr. Shin-ichi Muramatsu, Jichi Medical University, Japan.
3
Isolation and Culture of Rat Aortic Smooth Muscle Cells
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Primary rat SMC cultures were established according to a modification of the method of Ross, et al. [40 ]. Briefly, rat descending aorta was aseptically harvested, adherent fat and adventitia were removed and aortas were de-endothelialized via passage of an applicator. Aortic tissue was then minced and fragments were incubated (37 °C, 5 % CO2) in Dulbecco’s Modified Eagles Medium (DMEM, Life Technologies, Carlsbad, CA) for seven days to allow outgrowth.
Primary SMCs were cultured in T-75 tissue culture flasks (Thermo Scientific, Rochester, NY, USA) with supplemented DMEM. DMEM was supplemented with 10 % fetal calf serum (Life Technologies, Carlsbad, CA, USA), 1 % (v/v) antibiotic-antimycotic (Life Technologies, Carlsbad, CA, USA), and 1 % (v/v) 0.2 M L-glutamine (Lonza Walkersville, Walkersville, MD, USA). Media was stored at 4 °C for use up to 4 weeks. SMC were grown to 80 % or greater confluency and were passaged with trypsin-versene mixture (Lonza Walkersville, Walkersville, MD, USA) before use in experiments. Only cells between passages 3 and 8 were used.
Primary SMCs were cultured in T-75 tissue culture flasks (Thermo Scientific, Rochester, NY, USA) with supplemented DMEM. DMEM was supplemented with 10 % fetal calf serum (Life Technologies, Carlsbad, CA, USA), 1 % (v/v) antibiotic-antimycotic (Life Technologies, Carlsbad, CA, USA), and 1 % (v/v) 0.2 M L-glutamine (Lonza Walkersville, Walkersville, MD, USA). Media was stored at 4 °C for use up to 4 weeks. SMC were grown to 80 % or greater confluency and were passaged with trypsin-versene mixture (Lonza Walkersville, Walkersville, MD, USA) before use in experiments. Only cells between passages 3 and 8 were used.
4
Cytotoxicity Assays Reagents Purchase
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Trifluoroacetic acid (TFA), propidium iodide (PI), 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyletrazolium bromide (MTT), cis-platinum, taxol, all trans-retinoic acid (ATRA), and 10 hydroxyl-camptothecin (HCPT) were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Cell culture medium RPMI 1640 and fetal bovine serum (FBS) were purchased from GIBCO (Carlsbad, CA, USA). Penicillin-streptomycin and trypsin-versene mixture were purchased from BioWhittaker (Walkersville, MD, USA). Sterilized cell culture materials were purchased from Beckton Dickinson Labware (Franklin Labkes, NJ, USA).
5
Synthesis and Characterization of Iron Oxide Nanoparticles
6
Cell Culture of Human Cancer Lines
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Human cervical cancer HeLa and HEK293 cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA) and maintained in complete DMEM (Mediatech, Manassas, VA, USA), supplemented with 10% heat-inactivated fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA), and 1% penicillin and streptomycin (P/S; Lonza, Walkersville, MD, USA). Cells were grown as adherent cultures in a humidified atmosphere at 37°C in 5% CO2, sub-cultured after treatment with a trypsin-versene mixture (Lonza, Walkersville, MD, USA) for 2–5 min at room temperature, and washed and re-suspended in complete medium.
7
Cultivation of Huh7 and HepG2 Liver Cancer Cells
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Human hepatocellular carcinoma cells, Huh7 and HepG2, were purchased from ATCC (Old Town Manassas, VA, United States) and were maintained in Dulbecco’s-modified Eagle medium (DMEM; Gibco, Waltham, MA, United States) supplemented with 10% fetal bovine serum (FBS; Gibco, Waltham, MA, United States) and 1% penicillin/streptomycin (Invitrogen, Carlsbad, CA, United States) as described previously (Yin et al., 2021 (link)). Cell cultures were grown at 37°C and 5% CO2. Cells were passaged using trypsin-Versene mixture (Lonza, Walkersville, MD, United States), and resuspended in complete DMEM.
8
Culturing Human Cell Lines for Research
9
Culturing Human Cell Lines for Research
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Human embryonic kidney, HEK293, and human hepatocellular carcinoma, Huh7, cells were purchased from American Type Culture Collection (Manassas, VA), and HepaRG cells, terminally differentiated hepatic cells derived from a human hepatic progenitor cell line that retain many of the characteristics of primary human hepatocytes, were obtained from ThermoFisher Scientific, Waltham, MA. Huh7 cells, stably transfected with a firefly luciferase expression plasmid (Huh-FLuc) were generated as described previously [46 (link)]. Cells were maintained in complete Dulbecco Modified Eagle Medium (DMEM, Mediatech, Manassas,VA) supplemented with 10% heat-inactivated fetal bovine serum (FBS, Sigma-Aldrich, St. Louis, MO) and 1% penicillin and streptomycin (P/S, Lonza, Walkersville, MD). Cells were grown as adherent cultures in a humidified atmosphere at 37°C in 5% CO2, subcultured after treatment with trypsin-Versene mixture (Lonza, Walkersville, MD) for 2 to 5 min at room temperature, washed, and resuspended in complete DMEM.
10
Formulation and Characterization of Allantoin-Cyclodextrin Gels
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The materials used in this study were purchased and used as received: allantoin (Elton Chemical SA, Avlonas, Greece), β-cyclodextrin (β-CD) (Fluka, Ph. Eur.), hydroxypropyl-β-cyclodextrin (HP-β-CD) (Fluka, Ph. Eur.), Carbopol 934 or poly(acrylic acid), (PAA) (ServaFeinBiochemica, Heidelberg, Germany), triethanolamine (TEA) (Sigma-Aldrich, Schnelldorf, Germany), glycerol (CREMER OLEO, Hamburg, Germany), distilled water (FRX), NHDF (Normal Human Dermal Fibroblast) cell line from PromoCell, complete cell medium containing Eagle’s Minimal Essential Medium alpha, aMEM, Dulbecco’s Modified Eagle Medium without phenol red (DMEM), a mixture of penicillin-streptomycin-amphotericin B mixture of concentration of 1% (10 K/10 K/25 μg in 100 mL), EDTA (Trypsin-Versene mixture from Lonza), Fetal Bovine Serum (FBS) achieved from Biochrom GmbH (Berlin, Germany) and Phosphate Buffered Saline (PBS) from Invitrogen.
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