The human lung SCC cells H520, H226, the human pulmonary adenocarcinoma cells H358, H441, H460 and A549, the human esophageal SCC cells TE1, TE4, TE8, TE10 and human foreskin fibroblast HFF1 obtained from the American Type Culture Collection (Manassas, VA) and the normal human lung fibroblast cells NHLF obtained from Lonza (Portsmouth, NH) were grown in RPMI 1640 (H226, H358, H460, H520, TE1, TE4, TE8, TE10) or high glucose Dulbecco’s modified Eagle medium (H441, A549, HFF1, NHLF) supplemented with 10% heat-inactivated fetal bovine serum. Human umbilical vein endothelial cells (HUVEC) were purchased and grown in Endothelial Cells Growth Medium (Medium 200) supplemented with Low Serum Growth Supplement kit (Thermo Fisher Scientific, Rockford, IL). The lung squamous cell carcinoma cells EBC1, EBC2, SQ5, LK2 were kindly provided by Dr. Kiura Katsuyuki (Department of Respiratory Medicine, Okayama University Graduate School of Medicine and Dentistry, Okayama, Japan) and grown in RPMI 1640 supplemented with 10% heat-inactivated fetal bovine serum. All cell lines were cultured in 5% CO2 at 37°C.
Low serum growth supplement kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Low Serum Growth Supplement Kit is a laboratory product designed to support cell culture growth in low serum conditions. The kit contains a proprietary formulation of supplements that can be added to cell culture media to enhance cell proliferation and viability when serum levels are reduced.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Human umbilical vein endothelial cells (huvec), by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (2 mentions)
Medium 200, by Thermo Fisher Scientific (2 mentions)
Glacial acetic acid, by Merck Group (2 mentions)
Optima lc ms grade methanol, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Acetonitrile, by Thermo Fisher Scientific (2 mentions)
Hff 1, by American Type Culture Collection (1 mentions)
Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Exosome spin columns mw 3000, by Thermo Fisher Scientific (1 mentions)
Bodipy tr ceramide, by Thermo Fisher Scientific (1 mentions)
Prolong glass antifade mountant with nucblue stain, by Thermo Fisher Scientific (1 mentions)
A1 confocal microscope, by Nikon (1 mentions)
M200 medium, by Thermo Fisher Scientific (1 mentions)
Mda mb 231 cell, by American Type Culture Collection (1 mentions)
Tryple, by Thermo Fisher Scientific (1 mentions)
Mp468, by 3M (1 mentions)
17 protocols using low serum growth supplement kit
1
Lung and Esophageal Cancer Cell Line Cultivation
2
Exosomal Uptake by Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pooled HUVECs (Thermo Fisher Scientific) were cultured according to manufacturer’s instructions using Medium 200PRF completed with Low Serum Growth Supplement Kit (Thermo Fisher Scientific). Exosomes (25 μg/mL) derived from hMSCs grown onto TNs were stained with 10 μM BODIPY™ TR ceramide (Thermo Fisher Scientific) for 20 min at 37 °C. A mix without exosomes was used as the negative control. Excess unincorporated dye was removed from the labeled exosomes with Exosome Spin Columns (MW 3000) (Thermo Fisher Scientific), following the manufacturer’s protocol. The labeled exosomes or control solution were then incubated for 4 h with HUVECs seeded at a density of 5 × 104 cells/cm2 in 24-well plates the day before. After incubation, HUVECs were fixed with 4% paraformaldehyde solution in PBS for 10 min, then permeabilized for 10 min in 0.2% Triton X-100 (Sigma-Aldrich) prepared in PBS. After three washing cycles with PBS, the cells were incubated in 2% BSA solution in PBS for 1 h at RT. The cells were then stained with Alexa Fluor™ 488 Phalloidin (Thermo Fisher Scientific) for 1 h at room temperature. Nuclear staining was performed with ProLong™ Glass Antifade Mountant with NucBlue™ Stain (Thermo Fisher Scientific). The cells were observed with the Nikon A1 confocal microscope (Nikon, Minato, Tokyo, Japan) equipped with a 63X objective.
3
Tumor-Endothelial Cell Co-culture Model
4
Endothelial Cell Culture and Manipulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human umbilical vein endothelial cells (HUVECs; CC-2519; Lonza, MD, USA) were cultured in DMEM/F12 medium, supplemented with 10% FBS, EC growth supplement (50 μgmL−1; Lonza) and antibiotics, until confluency. Human aortic endothelial cells (HAECs; C-006-5C; Invitrogen) were cultured in medium 200 (M200500; Gibco) in the presence of low serum growth supplement kit (S003K; Thermo Fisher Scientific). Cells at passages four to seven were used for experiments. For adenoviral infection, HUVECs and HAECs were transfected with Ad-Vector or Ad-CMV-SOX4 (Vigene Biosciences) for 48 h. Morphology of HUVECs and HAECs were monitored by brightfield microscopy. For pharmacological experiments, HUVECs were incubated with metformin (200 μmolL−1, 72 h), IL-1β (10 ngml−1, 48 h) and TGF-β1 (10 ngml−1, 48 h) prior to Western blotting.
5
Diverse Cell Culture Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse embryonic stem cells were cultured in G-MEM BHK-21 (Gibco, 21710-025), 20% fetal bovine serum (FBS, Gibco, 10439-024), 1000 U/ml leukemia inhibitory factor (Chemicon, ESG1107), 0.1 mM non-essential amino acids (Gibco, 11140050), 0.1 mM β-mercaptoethanol and 10 mM sodium pyruvate (Gibco, 11360070) in a gelatin-coated flask. A mouse embryonic fibroblast cell line (Pasque et al., 2011 (link)) and a mouse myoblast cell line, C2C12 (ATCC, CRL-1772), were cultured in DMEM (Sigma, D5671) with 10% FBS (ThermoFisher Scientific, 26140079). Human lung stem cells were derived from fetal lung epithelial tips and grown as long-term self-renewing organoids (Nikolic et al., 2017 (link)). Human lymphoblast cell line K-562 (Merck, 89121407-1VL) was cultured in IMDM (ThermoFisher Scientific, 31980030) with 10% FBS. Human adult dermal fibroblasts (ThermoFisher Scientific, C0135C) were cultured in medium 106 (ThermoFisher Scientific, M106500) with a low serum growth supplement kit (ThermoFisher Scientific, S003K). Human neuroblastoma cell line SH-SY5Y (ATCC, CRL-2266) was cultured in Advanced DEME/F12 (Gibco,12634010) with 10% FBS (ThermoFisher Scientific, 26140079).
6
Glucocerebrosidase Lysosomal Activity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Eliglustat hemitartrate (EGT) (MCE, MedChem Express, NJ, USA), ambroxol hydrochloride (AMB) (Abcam, Cambridge, UK). Human anti-glucocerebrosidase (GBA) antibodies (GenTex), LAMP1, and LC3A/B antibodies (Cell signaling technology, Danvers, MA, USA). NuPAGE SDS running buffer, bolt 8% Bis-Tris Plus gel, Novex ECL chemiluminescent substrate reagents, sample reducing agents, media 106, low serum growth supplement kit, BCA protein assay kit (Thermo Fisher Scientific, Rockford, IL, USA). Sodium taurocholate hydrate, 4-Methylumbelliferyl β-D-glucopyranoside (Sigma-Aldrich, St. Louis, MO, USA), normocin (InvivoGen, San Diego, CA, USA).
7
T cell Transendothelial Migration Assay
8
Measuring Cytotoxicity and Inflammation in Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human aortic endothelial (HAE) cell and human colorectal adenocarcinoma HT-29
cell were purchased from American Type Culture Collection (ATCC, Manassas, VA,
USA). These two cells were used as well-established systems to measure LPS
cytotoxicity and gut-induced IL-8 secretion, respectively. The former was
maintained in cascade biologics medium 200 (Gibco Life technologies, Carlsbad,
CA, USA) containing low serum growth supplement kit (Gibco Life technologies),
and the latter in McCoy’s 5A medium (Gibco Life technologies) containing
10% fetal bovine serum and 1% penicillin-streptomycin solution (Gibco Life
technologies). These cells were cultured at 37°C in a humidified
incubator with 95% air and 5% CO2.
cell were purchased from American Type Culture Collection (ATCC, Manassas, VA,
USA). These two cells were used as well-established systems to measure LPS
cytotoxicity and gut-induced IL-8 secretion, respectively. The former was
maintained in cascade biologics medium 200 (Gibco Life technologies, Carlsbad,
CA, USA) containing low serum growth supplement kit (Gibco Life technologies),
and the latter in McCoy’s 5A medium (Gibco Life technologies) containing
10% fetal bovine serum and 1% penicillin-streptomycin solution (Gibco Life
technologies). These cells were cultured at 37°C in a humidified
incubator with 95% air and 5% CO2.
9
HUVEC Culture and Maintenance Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HUVECs were purchased from Lonza (Baltimore, MD, USA). Cells were cultured in M200 (Gibco, Grand Island, NY, USA) supplemented with an low serum growth supplement kit (Gibco) and incubated at 37°C in a humidified atmosphere with 5% CO2. The culture medium was replaced every 2 days until cells were confluent. All experiments were performed using cells at passages 3∼6.
10
Hepatic Esterase and Pancreatic Lipase Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Myricetin, TS, TC, BR, PL, NR, hepatic esterase from porcine (≥ 15 U mg−1 solid), and pancreatic lipase from porcine (Type II, 100–500 U mg−1 protein) were purchased from Sigma Aldrich Co. (St. Louis, MO, USA). Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum (FBS), and trypsin–EDTA were purchased from GE Healthcare (Chicago, Illinois, USA). Pilocarpine HCl and glycopyrrolate bromide were obtained from Tokyo Chemical Industry (Tokyo, Japan). Meditoxin, alfaxalone (Alfaxan), and xylazine HCl (Rompun) were purchased from Medy-Tox Inc. (Ochang, Korea), Careside (Gyeonggi-do, Korea), and Bayer Korea (Seoul, Korea), respectively. Antibiotic–antimycotic (10 ×), Medium 106, and low serum growth supplement kit were purchased from Gibco/Invitrogen (Grand Island, NY, USA), and AlamarBlue cell viability reagent was obtained from Thermo Scientific (Waltham, Massachusetts, USA). All other chemicals were of analytical and reagent grade.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!