Human pancreatic islets were cultured in Connaught Medical Research Laboratories (CMRL) 1066 medium, containing 5.5 mM or 20 mM glucose (where indicated) and supplemented with 10% FBS (Gibco), 110 U/mL penicillin (Gibco), 110 μg/mL streptomycin (Gibco), 50 μg/mL gentamicin (Gibco), 2 mM l -glutamine (GlutaMax, Gibco), and 1 mM sodium pyruvate (Gibco). Islet cell gentle dissociation was done using 0.05% Trypsin (Gibco) treatment. For bioluminescence recordings, 100 islets were plated to multiwell plates (LifeSystemDesign). For video time-lapse microscopy experiments, ∼20 human islets were plated to a 3.5-cm glass-bottom willco dish (WillCo Wells). For perifusion experiments on purified β-cells (Fig. 3A ), ∼25,000 FACS-sorted β-cells were attached to 35-mm dishes (Falcon). For the rest of the experiments, ∼50,000 dissociated islet cells were attached to 35-mm dishes (Falcon). All dishes were precoated with a homemade laminin-5–rich extracellular matrix derived from 804G cells, as described in ref. 54 (link).
35 mm dishes
Manufactured by BD
Sourced in United States
The 35-mm dishes are circular culture vessels commonly used in cell culture and tissue engineering applications. They provide a contained environment for the growth and maintenance of cells and tissues. The dishes are made of high-quality materials and are designed to meet the requirements of various laboratory protocols.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Trypsin, by Thermo Fisher Scientific (2 mentions)
Gentamicin, by Thermo Fisher Scientific (2 mentions)
Xfect transfection reagent, by Takara Bio (2 mentions)
Puromycin, by Thermo Fisher Scientific (2 mentions)
Px330, by Addgene (2 mentions)
Puc19, by Takara Bio (2 mentions)
Px330, by Takara Bio (2 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
L glutamine glutamax, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium dmem, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by GE Healthcare (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
15 protocols using 35 mm dishes
1
Culturing human pancreatic islets
2
Human Pancreatic Islet Isolation and Culture
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Human pancreatic islets were obtained from 4 different sources, summarized in Table 1 : (i) Prodo Laboratories company (ND and T2D islets); (ii) Alberta Diabetes Institute islet core center (UAL) (ND and T2D islets); and (iii) Islet Transplantation Center of Geneva University Hospital (ND islets). T2D donors had a history of T2D and/or HbA1c greater than 6.5%. Details of the islet donors are summarized in Table 1 . All procedures using human islets were approved by the ethical committee of Geneva University Hospital CCER 2017–00147. Human pancreatic islets were cultured in CMRL 1066 medium, containing 5.5 mM glucose and supplemented with 10% fetal bovine serum (Gibco), 110 U/ml penicillin (Gibco), 110 μg/ml streptomycin (Gibco), 50 μg/ml gentamicin (Gibco), 2 mM L-glutamine (GlutaMax, Gibco), and 1 mM sodium pyruvate (Gibco). Islet cell gentle dissociation was done using 0.05% Trypsin (Gibco) treatment. For lipidomic analysis, approximately 600 islets were plated to 35-mm dishes (Falcon). For bioluminescence recordings, 100 islets were plated to multi-well plates (LifeSystemDesign). For the rest of the experiments, approximately 50,000 dissociated islet cells were attached to 35-mm dishes (Falcon). All dishes were precoated with a homemade laminin-5-rich extracellular matrix derived from 804G cells as described in [92 (link)].
3
Human Neuroblastoma SH-SY5Y Cell Line Protocol
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Human neuroblastoma SH-SY5Y cells (passage 20) were a generous gift from Chia-Wen Tsai (China Medical University, Taichung, Taiwan). Cell culture methods refer to previously published literature [9 (link)]. DMEM, penicillin-streptomycin, trypsin-EDTA, and fetal bovine serum were purchased from Gibco, ThermoFisher Scientific (Waltham, MA, USA). Cells were plated on 35 mm dishes (BD Falcon, NY, USA) at a density of 1.0 × 106 cells per dish, and then incubated with 1 μM MK for 24 h and then exposed to 100 μM 6-OHDA for 18 h.
4
Dental Pulp Stem Cell Exposure to Polycaprolactone
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The study was approved by the Animal Care and Use Committee of the National Center for Geriatrics and Gerontology Research Institute and Aichi Medical University (permission #2016-5, 2017-25, issued on 25 March 2017). DPSCs were isolated according to a previous study [19 (link)] under relevant guidelines and regulations. Briefly, after extracting the upper canine teeth from 8- to 10-month-old beagle dogs (n = 3) (Kitayama Lab, Ina, Japan), DPSCs were isolated by enzymatic digestion and seeded on 35 mm dishes (Falcon, Corning, Tewksbury, MA, USA) in Dulbecco’s Modified Eagle’s Medium (DMEM) (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS; GE Healthcare UK Ltd., Amersham, UK). Cultured DPSCs (4th to 6th passage) at 70% confluence were subjected to various doses of PC (100, 200, 500 μM) (Sigma-Aldrich, St. Louis, MO, USA) for the time points of 24, 48, and 72 h.
5
Bone Marrow Hematopoietic Assay
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Single cell suspensions were prepared from bone marrow and mixed with methylcellulose-containing medium with cytokines (Casanova-Acebes et al., 2013 (link)). Cells (5-7.5 × 104) were plated in duplicate 35 mm dishes (Falcon, BD) and incubated under 20% O2 and 5% CO2 in a water-jacketed incubator. Hematopoietic colonies (CFU-Cs) were scored after 6-7 days in culture.
6
Neuroblastoma Cell Migration Assay
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Neuroblastoma SH-SY5Y-trkB and NMB-7 cells were grown in 35 mm dishes (Falcon) and cultured in RPMI 1640 medium (Life Technologies) supplemented with 5% fetal bovine serum. 3F8 (50 nM), 3F8+PP2(10 μM), SS58 (10 μM) and SS58+PP2 (10 μM) were added and time-lapse photographs were taken at 100X magnification using a Nikon camera and a microscope adapted with a 37°C chamber.
7
Circadian Rhythm Monitoring in Fibroblasts
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Per2::Luc KI MEFs and mutant NIH3T3 fibroblasts were plated in 35-mm dishes (Falcon, United States) at a 1.5 × 105 cell density. After 24 h of incubation, the cells were transfected with selected miRNAs (50 nM) using Lipofectamine 3000 according to the manufacturer’s protocol and incubated for 24 h (Invitrogen). The cells were synchronized by 200 nM DEX treatment for 2 h, and the bioluminescence was recorded for 5 days with a real-time bioluminescence device (Kronos Dio) in recording medium with 100 µM d -luciferin (Promega). Light emissions were integrated for 1 min at intervals of 10 min using a dish-type wheeled luminometer. The raw data presented were normalized based on the first minimum bioluminescence point and detrended bioluminescence time series data were obtained by subtracting the 12-h moving average from the luminescence intensity with the background eliminated.
8
Huh7 Cells Exposed to HCCMF RF EMF
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Huh7 cells – seeded into six-well plates or 35 mm dishes (Falcon; Corning) – before receiving either AM RF EMF treatment. 300,000 cells were left to adhere overnight and cultured in Dulbecco's Modified Eagle's Medium (DMEM; Cellgro) supplemented with heat-inactivated foetal bovine serum (FBS, final concentration 10%; Atlanta Biologicals). Cells were exposed to either HCCMF (hepatocellular carcinoma specific AM RF EMF) or no treatment for 30 min, 1 h, 3 h, or 6 h. 30 min before completion of treatment Fluo-4 stain mix was added directly to each well/dish (Fluo-4 Calcium Imaging Kit; Molecular Probes). Dishes/plate were placed on a rocker at room temperature for 5 min of gentle distribution of Fluo-4 stain. Cells were then placed back into the 37 °C incubator and exposed for 30 min to HCCMF (completing the required time of HCCMF exposure). Following completion of treatment exposure, the dishes/plates were placed at room temperature for 30 min (in the dark). Cells were then washed using 1·5 mL of LCIS (Live Cell Imaging Solution; Molecular Probes) then placed in two millilitres of fresh LCIS and fluorescent intensity was read using a Fluostar fluorescent plate reader (BMG LABTECH) (485 excitation/520 emission). All experiments were performed at least twice with representative experiments shown. Statistics: one-way t-test was utilized.
9
Hesperetin Inhibits HCASMC Contraction
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HCASMCs were cultured in 35-mm dishes (BD Falcon, NY) at a density of 1 × 105 cells/dish. When the cell confluence reached 90%–100%, the medium was replaced with FBS-free and growth factor-free HuMedia-SB2 to obtain the hypercontractile type of HCASMCs. After treatment with HuMedia-SB2 for 24 hours, the cells were pretreated with hesperetin for 30 minutes at 37°C. Next, 30 μM SPC was added to the medium, and time-lapse recording of HCASMC contraction was performed using an all-in-one fluorescence microscope BZ-9000 (Keyence, Osaka, Japan).
10
Erythroid Differentiation of Human CD34+ Cells
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HUDEP-2 cells and HiDEP-1 cells were grown in Stemline II Hematopoietic Stem Cell Expansion Medium (Sigma) supplemented with 50 ng/mL recombinant stem cell factor (SCF; R&D systems), 3 IU/mL recombinant erythropoietin (EPO; Kyowa Kirin), 1 μM dexamethasone (Dex; Sigma), and 1 μg/mL doxycycline (Sigma). A total of 1 × 105 human CD34+ progenitor cells derived from bone marrow or peripheral blood (Lonza) were cultured on 35-mm dishes (BD Falcon) in StemSpan SFEMII medium with 50 ng/mL SCF, 5 IU/mL EPO, and 10 ng /mL interleukin 3 (IL-3; R&D systems). The medium was changed on day 4 to SCF, EPO, and 1 μM Dex, and the cells were transferred to 60-mm dishes (BD Falcon). Seven days after the onset of differentiation, cells were counted and 1 × 106 cells were re-plated on 100-mm dishes (BD Falcon) with SCF, EPO, and Dex. The rat myoblast cell line H9c2, HEK293T, and PLAT-E cells were maintained in Dulbecco’s modified Eagle’s medium (Thermo Fisher Scientific) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and penicillin/streptomycin. All cells were cultured at 37 °C under 5% CO2.
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