In order to screen lysoPC‐sensitive lung epithelial cells, the cell proliferation of large cell human lung carcinoma cells (NCI‐H460 and NCI‐H661) that expresses easily p53 mRNA detection, non‐SCLC cells (NCI‐H1299), lung ADC (A549 and NCI‐H1650), SCLC cells (NCI‐H1688), gene‐edited lung ADC epithelia (A549p53−), lung bronchial ADC epithelia (SPC‐A1 cells) and normal bronchial epithelia (HBE135‐E6E7) were measured using CCK8 (C0037, Beyotime, Shanghai, China) at 3, 6, 12, 24 and 48 h after treatment with exogenous lysoPC (L1381, Sigma‐Aldrich, MO, USA) at concentrations of 25, 50, 100 or 200 μM. Cells were cultured in full media consisting of RMPI 1640 (KGM31800‐500, KeyGEN, Jiangsu, China) containing 10% fetal bovine serum (F8318, Sigma‐Aldrich, MO, USA), penicillin 100 U/ml and streptomycin 100 mg/ml and in the cell incubator at 37°C with 5% CO2.
F8318
Manufactured by Merck Group
Sourced in United States
The F8318 is a piece of laboratory equipment manufactured by the Merck Group. It is a versatile and reliable instrument designed for various laboratory applications. The core function of the F8318 is to perform precise and consistent measurements, ensuring accurate data collection and analysis. Further details about the intended use or specific applications of this product are not available.
Automatically generated - may contain errors
Lab products found in correlation
L glutamine, by Merck Group (2 mentions)
Penicillin streptomycin, by Merck Group (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
L1381, by Merck Group (1 mentions)
Smart silencer nc, by RiboBio (1 mentions)
M0512, by Merck Group (1 mentions)
Cell incubator, by Thermo Fisher Scientific (1 mentions)
Cell strainer, by Biosharp (1 mentions)
D0821, by Merck Group (1 mentions)
Hek293t, by American Type Culture Collection (1 mentions)
Exfect transfection reagent, by Vazyme (1 mentions)
A4192101, by Thermo Fisher Scientific (1 mentions)
9 protocols using f8318
1
Lysopc-Sensitivity Screening of Lung Cells
2
Neonatal Rat Cardiomyocyte Inflammatory Response
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Neonatal rat cardiomyocytes (NRCM) were obtained according to the previous method (Zhang et al., 2018 (link)). Different concentrations of PRP (1%,5%, and 10%) and inhibitors (AKTi and Rapamycin, MCE, HY-10355, HY-10219) were added into the medium containing 10% fetal bovine serum (Sigma, F8318) and 1% penicillin-streptomycin solution (Hyclone, SV30010), and the same amount of PBS was added to the control group. After 2 h of incubation in a 37°C, incubator, LPS (1 μmol/L) was added for 24 h (Figure1 ).
3
Enteric Glial Cell Knockdown Study
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Rat enteric glial cells (HZ-CRL-2690, ATCC, USA) were cultured in Dulbecco's modified Eagle's medium (DMEM; 11965092, Gibco, USA) containing 10% fetal bovine serum (FBS; F8318, Sigma-Aldrich, USA) and 1% penicillin and streptomycin (15140-122, Gibco, USA). The cells were maintained at 37 °C with 5% CO2 under saturated humidity. Rat H9c2 cells were used as a positive reference to detect the basal expression of LOC108352929 and Phf11. Smart silencer technology was employed to knock down the expression of lncRNA LOC108352929 to observe its effect on the expression of Phf11, TNF-α, and IL-1β in EGCs. EGCs were categorized into four groups: Smart Silencer-LOC108352929 group (SS), Smart silencer-NC group (NC), transfected reagent control group (mock), and unprocessed cell control group (blank). Two replicate wells were used for each group. Smart silencer-LOC108352929 and Smart silencer-NC were custom-synthesized by RIBOBIO (China). After 48 h of culture, cells were collected for RNA extraction.
4
Methylcellulose-based 3D Spheroid Assay
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Spheroid formation assay was performed essentially as previously described [43 (link)]. For spheroid generation we used 20% of the methylcellulose stock solution (M0512, Sigma-Aldrich, St. Louis, MO, USA) and 80% culture medium corresponding to final 0.24% methylcellulose. The culture medium for all cells were used by DMEM-F12 supplemented with 10% FBS (F8318, Sigma-Aldrich, St. Louis, MO, USA). Low attachment 24-well plates (3473, Corning, NY, USA) at a density of 5000 cells/well. After 10 days of culture, the spheroids with diameter over 200 μm were counted and photographed.
5
Cell Culture Protocols for Microglial, Neuronal, and Endothelial Cells
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The microglial cell line (BV2, BNCC337749, Beina Biology, Beijing, China) and hippocampal neuronal cell line (HT22, BNCC337749, Beina Biology, Beijing, China) were cultured in high glucose Dulbecco’s modified Eagle’s medium (DMEM/H, C11995500BT, Gibco) supplemented with 10% (v/v) fetal bovine serum (FBS, F8318, Sigma) and 1% penicillin/streptomycin and were passaged every 2 to 3 days. Mouse brain microvascular endothelial cells (bEnd.3, CC-Y2019, Shanghai Jining Industrial LTD, Shanghai, China) were cultured in DMEM/H with 15% (v/v) FBS and 1% penicillin/streptomycin; the culture medium was changed every 2 days, and the cells were passaged every 3 to 4 days. bEnd.3 cells below passage 9 were used in the experiments. The three types of cells were cultured at 37 °C in a humidified 5% CO2 atmosphere.
6
In vitro Culture of Mouse Conjunctival Goblet Cells
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In this study, we chose mouse conjunctival GCs for in vitro experiments, conjunctival GCs were dissociated and cultured using a protocol published by García-Posadas et al. (14 (link)). The cultured cells were diluted to 1×106 cells/mL, and evenly added to 24-well plates. These cells were randomly divided into control group, carbomer group, and VAP group, VAP gel and carbomer gel were added into VAP and carbomer group respectively, then fetal bovine serum (FBS, 10%, F8318, Sigma-Aldrich, St. Louis, MO, USA), 2 mM L-glutamine (G7513, Merck, Darmstadt, Germany), 100 µg/mL penicillin-streptomycin (V900929, Merck) were added to 24-well plates.The above cells were cultured in cell incubator (51032124, ThermoFisher Scientific) at 37 °C and 5% CO2 for another 24 h.
7
Isolation of Mouse Cortical Neurons
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Newborn mice aged 1–3 days were subjected to gentle meningeal dissection using ophthalmic forceps, scissors, and other tools. Subsequently, the brain tissue was carefully peeled off and placed in a 0.25% trypsin solution (Gibco, 15050065). The tissue was then incubated in a cell culture incubator at 37 °C for 5 min. After the incubation period, the trypsin reaction was stopped by adding complete culture medium, and the cell suspension was filtered through a cell strainer (Biosharp). The filtrate was collected and subjected to centrifugation at 1000g for 10 min. The supernatant was discarded, and the cells were resuspended in complete culture medium. The cells were then seeded in T-75 cell culture flasks containing 10% fetal bovine serum (FBS) (Sigma, F8318)-F12/DMEM culture medium and were subsequently maintained through medium changes and passaging for experimental use.
8
HEK293T Cell Transfection Protocol
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HEK293T (ATCC, Manassas, VA) were cultured in Dulbecco’s modified Eagle medium (DMEM; D0821, Sigma Aldrich) containing 10% fetal bovine serum (FBS; F8318, Sigma Aldrich) at 37°C in a humidified cell incubator with 5% CO2 and 95% O2. The plasmids were transfected into the HEK293 T cells using ExFect Transfection Reagent (T101-01/02, Vazyme) according to the manufacturer’s instructions.
9
Culturing Human Thyroid Cell Lines
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Human thyroid epithelial (Nthy-ori3-1) cells and PTC cell lines (TPC-1 and IHH-4 cells) were ordered from COBIOER (CBP61205, CPB60257, CPB61201, Nanjing, China, http://www.cobioer.com/). PTC cell line (MDA-T68 cells) was obtained from ATCC (CRL-3353, Manassas, VA, USA). Nthy-ori3-1 cells were cultured in RPMI-1640 medium (22400105, ThermoFisher, Walthma, MA, USA) which was supplemented with 10% fetal bovine serum (FBS, F8318, Sigma-Aldrich, St. Louis, MO, USA). TPC-1 cells were cultured in RPMI-1640 containing 10% FBS and 100 U/mL streptomycin/ penicillin (V900929, Sigma-Aldrich, USA). IHH-4 cells were cultured in a mixed medium comprising DMEM (A4192101, ThermoFisher, USA) and RPMI-1640 at a ratio of 1:1 and supplemented with 10% FBS. MDA-T68 cells were cultured in RPMI-1640 containing 10% FBS and 2 mM L-glutamine (G7513, Sigma-Aldrich, USA). All cells were cultured at 37°C with 5% CO 2 .
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