The obtained tissues (tumor or peri-tumor) were immersed in Dulbecco's modified Eagle's medium (DMEM; HyClone; GE Healthcare Life Sciences) supplemented with 2% penicillin-streptomycin (HyClone; GE Healthcare Life Sciences) for at least 30 min, and then washed with PBS (HyClone; GE Healthcare Life Sciences) three times. The tissues were minced with scalpels into 1.0×1.0×1.0 mm3 fragments and tiled on the bottom of a culture bottle. The cells were cultured in DMEM supplemented 20% FBS (HyClone; GE Healthcare Life Sciences) and 1% penicillin-streptomycin. After 3 h, the culture bottle was turned over. Cultures were incubated at 37°C in a humidified atmosphere of 5% CO2 and cell growth was observed under an inverted microscope (magnification, ×100). Upon reaching 90% confluence, the areas which epithelioid cells had grown in clusters were marked on the culture bottle when the cells were observed under the phase-contrast microscope (magnification, ×100), and according to the markers, epithelioid cells were wiped with a cotton stick dipped in 10 µl of trypsin. The remaining cells were digested with 0.25% trypsinase for 1 min; digestion was terminated with 3 ml of medium containing serum, and cells were sub-cultured into fresh bottles. Cells at passage (P)3-P5 were used in subsequent experiments.
Hyclone
Manufactured by GE Healthcare
Sourced in United States, United Kingdom, China, Australia, Germany, Sweden, Austria, Israel, Japan, New Zealand, Cameroon, France
HyClone is a line of laboratory equipment and consumables manufactured by GE Healthcare. The core function of HyClone products is to provide reliable and consistent solutions for cell culture and bioprocessing applications in research and clinical settings. HyClone offers a range of products, including media, sera, buffers, and other cell culture-related supplies.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (208 mentions)
Streptomycin, by Thermo Fisher Scientific (113 mentions)
Penicillin, by Thermo Fisher Scientific (104 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (89 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (83 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (69 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (37 mentions)
Rpmi 1640, by Thermo Fisher Scientific (30 mentions)
Fetal bovine serum (fbs), by GE Healthcare (26 mentions)
Streptomycin, by Merck Group (21 mentions)
L glutamine, by Thermo Fisher Scientific (21 mentions)
Streptomycin, by GE Healthcare (20 mentions)
Mda mb 231, by American Type Culture Collection (20 mentions)
Glutamax, by Thermo Fisher Scientific (19 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (17 mentions)
Penicillin, by GE Healthcare (16 mentions)
Penicillin streptomycin, by GE Healthcare (16 mentions)
Penicillin, by Merck Group (14 mentions)
Dulbecco modified eagle medium (dmem), by GE Healthcare (12 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (12 mentions)
1 189 protocols using hyclone
1
Efficient Primary Tumor Cell Culture
2
Protective Effects of Ginsenoside Rg3 on Acute Pancreatitis
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Rat pancreatic acinar AR42J cells were purchased from the American Type Culture Collection (ATCC). The cells were cultured in F12K medium (HyClone; GE Healthcare Life Sciences) supplemented with 20% FBS (HyClone; GE Healthcare Life Sciences), 100 U/ml penicillin (HyClone; GE Healthcare Life Sciences) and 100 mg/ml streptomycin (HyClone; GE Healthcare Life Sciences) in a humidified atmosphere containing 5% CO2 at 37°C. The AR42J cells were pre-incubated with or without Rg3 (cat. no. HY-N0603; MedChemExpress) at 37°C for 1 h and then treated with cerulein (Cn; 10−8 M; cat. no. HY-A0190; MedChemExpress) for a further 24 h to examine the protective effects of ginsenoside Rg3 on AP.
3
Culturing Gastric Cancer Cell Lines
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The human gastric cancer cell lines AGS and NCI-N87 were purchased from the American Type Culture Collection (ATCC). The normal human gastric epithelium cell line (GES-1) was purchased from Procell Life Science & Technology Co., Ltd. (CL-0563). Cell authenticity was identified using STR files. Cells were cultured at 37°C with Dulbecco's modified Eagle's medium (DMEM) (HyClone; GE Healthcare Life Sciences) supplemented with 10% fetal bovine serum (HyClone; GE Healthcare Life Sciences), 100 U/ml penicillin (HyClone; GE Healthcare Life Sciences) and 100 g/ml streptomycin (HyClone; GE Healthcare Life Sciences) in a humidified atmosphere containing 5% CO2.
4
Culturing Human Colon Cancer Cell Lines
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The human colon cancer cell lines HT-29, Caco-2 and HCT-116 were obtained from the State Key Laboratory of Molecular Oncology, National Cancer Center/Cancer Hospital, Chinese Academy of Medical Sciences (cell line not authenticated). HT-29, Caco-2 and HCT-116 cells were maintained in Dulbecco's modified Eagle's medium: Nutrient mixture F-12 media (DMEM/F-12; HyClone; GE Healthcare Life Sciences), minimum essential medium/Earle's balanced salt solution (MEM/EBSS; HyClone; GE Healthcare Life Sciences) and Iscove's modified Dulbecco's medium (IMDM; HyClone; GE Healthcare Life Sciences), respectively. They were supplemented with 10% fetal bovine serum (FBS; HyClone; GE Healthcare Life Sciences) and 1% penicillin/streptomycin (HyClone; GE Healthcare Life Sciences) at 37°C in a humidified incubator containing 5% CO2. Caco-2 cells were co-treated with 1% non-essential amino acids (HyClone; GE Healthcare Life Sciences).
5
Isolation and Culture of Mouse and Rat Immune Cells
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The mouse brain endothelial cell line bEnd.3 (ATCC CRL-2299) was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and the mouse macrophage cell line, RAW 264.7, kindly provided from by Professor Namhyun Jung from Korea University. All cells were cultured in Dulbecco’s modified eagle medium (DMEM; Welgene, Daegu, Korea) supplemented with 10% fetal bovine serum (FBS; HyClone, GE Healthcare Life Sciences, Logan, UT, USA) and 1% penicillin/streptomycin (HyClone, GE Healthcare Life Sciences, Logan, UT, USA) at 37 °C in a 5% CO2 humidified atmosphere.
Rat peripheral blood mononuclear cells were isolated from heparinized blood obtained from normal Sprague–Dawley (SD) rats via density gradient centrifugation (Ficoll-Plaque Plus, GE Health Care Life Sciences, Pittsburgh, PA, USA) according to the manufacturer’s instruction. Differentiation into macrophages was induced by a recombinant rat macrophage colony-stimulating factor (15 ng/mL; Peprotech, Rocky Hill, NJ, USA) with 1% penicillin/streptomycin (HyClone, GE Healthcare Life Sciences, Logan, UT, USA) and 10% heat-inactivated FBS (HyClone, GE Healthcare Life Sciences, Logan, UT, USA) for 7 days.
Rat peripheral blood mononuclear cells were isolated from heparinized blood obtained from normal Sprague–Dawley (SD) rats via density gradient centrifugation (Ficoll-Plaque Plus, GE Health Care Life Sciences, Pittsburgh, PA, USA) according to the manufacturer’s instruction. Differentiation into macrophages was induced by a recombinant rat macrophage colony-stimulating factor (15 ng/mL; Peprotech, Rocky Hill, NJ, USA) with 1% penicillin/streptomycin (HyClone, GE Healthcare Life Sciences, Logan, UT, USA) and 10% heat-inactivated FBS (HyClone, GE Healthcare Life Sciences, Logan, UT, USA) for 7 days.
6
Culturing HaCaT Cells in DMEM with FBS
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HaCaT cells (GCC-AO0003CS; Shanghai Jikai Gene Medical Technology Co., Ltd.) were cultured in DMEM (HyClone; GE Healthcare Life Sciences) supplemented with 10% heat-inactivated FBS (Gibco; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin (HyClone; GE Healthcare Life Sciences), and 100 µg/ml streptomycin (HyClone; GE Healthcare Life Sciences). The HaCaT cell line was authenticated by the supplier using STR profiling. The cells were maintained in a humidified incubator at 37°C with 5% CO2.
7
Cultivation of Urothelial Cell Lines
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The human urothelial cell lines RT4 and T24 were purchased from the American Type Culture Collection (ATCC® HTB-2™, ATCC® HTB-4™ and ATCC® CRL-1573™, respectively). RT4 and T24 cells were cultured in DMEM (HyClone; GE Healthcare Life Sciences, USA) containing 1% penicillin-streptomycin solution (HyClone; GE Healthcare Life Sciences, USA) and 10% fetal bovine serum (FBS, HyClone; GE Healthcare Life Sciences, USA) at 37°C in a humidified incubator with 5% CO2.
8
Production and Purification of Recombinant Antibodies
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As described previously (7 (link)), human embryonic kidney fibroblast 293T cells (HEK-293T, ATCC CRL 11268TM) were split into culture dishes (100 × 20 mm) in high glucose DMEM (HyClone, GE Healthcare), supplemented with heat inactivated fetal bovine serum (FBS, 10%), glutamine (1%) and penicillin-streptomycin (1%, HyClone, GE Healthcare). After 16–24 h, at 70–80% confluence, the medium was replaced with SFM4Transfx-293 (HyClone, GE Healthcare) supplemented with 10% FBS, 1% glutamine and 1% penicillin-streptomycin (HyClone, GE Healthcare). Cells were co-transfected 8 h later, using Lipofectamine LTX Reagent (Invitrogen), with IgH and IgL encoding plasmids (5 μg). Supernatants were collected after 7 days, and the antibodies were purified on protein G-sepharose beads (GE Healthcare, Life Technologies). Glycine buffer (0.1 M, pH 3.5) and Tris-HCl (1 M, pH 8) were used for antibody elution and pH neutralization, respectively. Purified antibody was dialyzed in PBS and its concentration was determined by Nanodrop. Antibody integrity was assessed on NuPAGE 4–12% BisTris gels (Invitrogen) stained with Coomassie blue.
9
Osteoblast Differentiation and Mechanistic Studies
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MC3T3-E1 mouse calvarial pre-osteoblasts were purchased from The Cell Resource Center of the Shanghai Institutes for Biological Sciences of The Chinese Academy of Sciences (Shanghai, China), and were cultured in α-Minimum Essential Medium (α-MEM; HyClone; GE Healthcare Life Sciences, Logan, UT, USA) supplemented with 10% fetal bovine serum (FBS; Biological Industries, Kibbutz Beit Haemek, Israel), 100 µg/ml streptomycin and 100 U/ml penicillin (HyClone; GE Healthcare Life Sciences). Cells were maintained in a cell culture incubator with 5% CO2 at 37°C. The medium was replaced every other day. Cells at 80% confluence were reseeded into tissue culture flasks following treatment with 0.25% trypsin (HyClone; GE Healthcare Life Sciences) for 1–2 min at 37°C. For osteoblastic differentiation experiments, cells were treated with osteogenic supplement containing 50 µg/ml L-ascorbic acid (Sigma-Aldrich; Merck KGaA) and 10 mM β-glycerophosphate disodium salt hydrate (Sigma-Aldrich; Merck KGaA) for 9 days at 37°C. For mechanistic studies, MC3T3-E1 cells were pretreated with DKK1 (0.5 µg/ml) for 6 h at 37°C, and were then treated with arbutin (100 µM) for 3 days at 37°C.
10
Isolation and Labeling of Mouse Bone Marrow Stromal Cells
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Bone marrow (BM) cell suspension was obtained by flushing marrow cavity of four- to five-week-old male mice with α-minimum essential medium (α-MEM; HyClone; GE Healthcare Life Sciences, Logan, UT, USA). Then, the BM cells were cultured with α-MEM containing 10% heat-inactivated fetal bovine serum (FBS; HyClone; GE Healthcare Life Sciences) and penicillin-streptomycin (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany). Cells were cultured at 37°C in a 5% CO2 humidified incubator. To acquire putative BMSCs, non-adherent cells and tissue debris were removed by phosphate-buffered saline (PBS) washing after 72 h (13 (link)). The culture medium was changed every 3 days until the cell density reached about 90%. BMSCs were detached by 0.25% trypsin containing 0.02% EDTA (HyClone; GE Healthcare Life Sciences) and expanded. All BMSCs were utilized for subsequent experiments.
To better observe the BMSCs, cells were labeled by cell tracker. The methods were as follows: BMSCs were suspended at a density of 3×106/ml with α-MEM. Every 1×106 cells were incubated with 3 µl CM-Dil (1 µg/ml) (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA) at 37°C for 5 min then 4°C for 15 min. Cells were washed twice and suspended by α-MEM.
To better observe the BMSCs, cells were labeled by cell tracker. The methods were as follows: BMSCs were suspended at a density of 3×106/ml with α-MEM. Every 1×106 cells were incubated with 3 µl CM-Dil (1 µg/ml) (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA) at 37°C for 5 min then 4°C for 15 min. Cells were washed twice and suspended by α-MEM.
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