Human kidney glomerular endothelial cells (HKGECs) were cultured in endothelial cell media (ScienCell; Carlsbad, CA, USA) containing vascular endothelial growth factor (VEGF; Sigma-Aldrich; St. Louis, MO, USA; 2.5–5 μg/ml) in basal medium (ScienCell), 5 % fetal bovine serum (FBS; ScienCell), 1 % endothelial cell growth supplement (ECGS; ScienCell) and 1 % penicillin/streptomycin (P/S; ScienCell). Cells were maintained at 37 °C with 5 % CO2. For treatment, HKGECs were cultured for 24 h at low density in flasks or plates pre-coated with fibronectin and washed in PBS. Cells were treated with TGF-β1 (10 ng/ml or 20 ng/ml; Sigma-Aldrich) alone, TGF-β1 plus 10 μM GM6001 (Calbiochem; Darmstadt, Germany), TGF-β1 plus the MMP-9 inhibitor I (Merck Chemicals; Darmstadt, Germany) at different dosages (0.05 nmol/ml, 0.25 nmol/ml, and 0.5 nmol/ml), recombinant MMP-9 (rMMP-9; 2 μg/ml; Biomol International; Plymouth Meeting, PA, USA), or rMMP-9 (2 μg/ml) plus the gamma secretase inhibitor (GSI; 5–10 μM; Merck Millipore; Billerica, MA, USA). Our study does not require any human or animal ethics approval.
Endothelial cell media (ecm)
Manufactured by ScienCell
Sourced in United States
Endothelial cell media is a specialized cell culture medium designed to support the growth and maintenance of endothelial cells. It provides the necessary nutrients and growth factors required for the optimal proliferation and differentiation of endothelial cells in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by ScienCell (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Huvecs, by ScienCell (2 mentions)
Mmp 9 inhibitor 1, by Merck Group (1 mentions)
Vascular endothelial growth factor (vegf), by Merck Group (1 mentions)
Tgf β1, by Merck Group (1 mentions)
Endothelial cell growth supplement (ecgs), by ScienCell (1 mentions)
1640 medium, by Thermo Fisher Scientific (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Sonic dismembrator model 100, by Thermo Fisher Scientific (1 mentions)
Zetasizer nano zs90 size analyzer, by Malvern Panalytical (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Facsaria sorter, by BD (1 mentions)
Human umbilical vein endothelial cells (huvec), by RIKEN BioResource Center (1 mentions)
Fibronectin, by ScienCell (1 mentions)
Anti pstat3 tyr705, by Cell Signaling Technology (1 mentions)
Anti jnk, by Abcam (1 mentions)
Anti ki67, by Abcam (1 mentions)
Anti β actin, by Abcam (1 mentions)
Anti nf κb, by Abcam (1 mentions)
16 protocols using endothelial cell media (ecm)
1
Modulation of Human Kidney Glomerular Endothelial Cells
2
Characterization of YTHDF3 Knockout SiHa Cell Line
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HFF-1 (Human epithelial foreskin fibroblast cells), SiHa, and CaSki cell lines were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Human lymphatic endothelial cells (HLECs) were maintained by endothelial cell media from ScienCell (CA) and were obtained from iCell Bioscience Inc (Shanghai, China). The SiHa and CaSki cells were incubated in 1640 medium (Gibco, USA), meanwhile the HFF-1 cell line was grown in DMEM medium (Gibco, USA). All of the medium was augmented with 10% fetal bovine serum (Excell, Uruguay), 100 U/mL penicillin, and 100 U/mL streptomycin (BI, USA). Cells were incubated in a humidified incubator at 37°C with 5% CO2. The short tandem repeat (STR) genotyping was confirmed for these cell lines. Furthermore, Haixing Biosciences (Guangzhou, China) used CRISPR-Cas9 technology to construct knockout YTHDF3 SiHa cells (YTHDF3-/- SiHa). The specific sgRNAs were listed in the Supplementary Table 2
3
Isolation and Characterization of HUVECs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
This study was authorized by the Research Ethics Committee of Universiti Kebangsaan Malaysia Medical Centre (Study Code: UKM PPI/111/8/JEP-2019-671). Umbilical cords were obtained from consenting donors at the labor room of Hospital Canselor Tuanku Mukhriz, Malaysia. Informed consent was obtained from all the donors. HUVEC were derived from fresh human umbilical cords using collagenase digestion (Gibco-Invitrogen Corp., Grand Island, NY, USA) as previously reported [26 (link)]. HUVEC were identified by the characteristic cobblestone morphology and expression of CD31 and vonWillebrand factor by immunocytochemistry. The cells were cultivated in endothelial cell media (ScienCell Research Laboratories, San Diego, CA, USA) at 37 °C in a humidified environment containing 5% CO2 and 95% air. The medium was replaced every two days. For subsequent experiments, HUVEC were subcultured at 80% confluency until they reached passage three.
4
Functionalization of TiO2 Nanoparticles for Biological Studies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Titanium dioxide nanoparticles (TiO2 NPs) were first prepared by dispersing 5 mg of p25 TiO2 nanopowder (MR: 79.87) with primary size of 21 nm (Sigma-Aldrich, St. Louis, MO, USA) in 1 mL of water to give 62.5 mM concentration and sonicated using Sonic Dismembrator model 100 (Thermo Fisher Scientific, Waltham, MA, USA) for 45 s. Subsequently, the suspension was further diluted with endothelial cell media (ScienCell Research Laboratories, CA, USA) into either 100 µM or 500 µM and sonicated for another 45 s. Hydrodynamic size and surface zeta potential of the NPs were then measured using Zetasizer Nano ZS90 size analyzer (Malvern Panalytical, Malvern, England). FITC was covalently tagged onto TiO2 NPs according to the previous study [16 (link)].
5
Glioblastoma Cell Line Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human glioblastoma cell lines LN229 and U87MG were obtained from the ATCC. LN229‐Luc2 cells were derived from a stable transfection of pLuc2‐iRFP and selected using a BD FACSAria sorter (BD Biosciences). LN229‐Luc2 cells were stably transfected with shRNA against PSMB8 and selected by puromycin. The cells were grown in RPMI‐1640 containing 10% FBS (Life Technologies) in a humidified atmosphere of 5% CO2 at 37°C. HUVEC were purchased from the Bioresource Collection and Research Center in Taiwan and cultured in endothelial cell media (ScienCell Research Laboratories).
6
Renal Glomerular Endothelial Cell Response to LPS and Shiga Toxin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human renal glomerular endothelial cells (RGECs) were cultured in endothelial cell media (ScienCell Research Laboratories, Carlsbad, CA) at 37 °C with 5% CO2. Briefly, 96-well plates were precoated with 7.5 μg/mL fibronectin (ScienCell Research Laboratories, Carlsbad, CA) for two hours at room temperature and washed once with phosphate-buffered saline without calcium and magnesium. Next, RGECs were seeded into 96-well plates at a density of 4 × 104 cells/well. After 24 h of incubation, the cells were treated with 30 μg/mL lipopolysaccharide (LPS) and/or Shiga toxin 2 (Stx-2) (0.1, 1.0, or 10 ng/mL) (Nacalai Tesque, Kyoto, Japan) for 48 h. Subsequently, culture supernatants were collected, and stored at −80 °C until further processing.
7
Angiogenesis Modulation in Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Recombinant human SF20 protein (Cat No. ab86915) and anti-SF20 antibody (Cat No. ab187919) were purchased from Abcam. Recombinant murine SF20 (Cat No. 210-25), human FGF (Cat No. 100-18B), human EGF (Cat No. GMP100-15), M-CSF (Cat No. 315-02) were acquired from PeproTech. Endothelial cell media (Cat No. 1001) was purchased from ScienCell. Mouse IL-6 (Cat No. 1210602), MCP-1 (Cat No. 1217392) and TNF-α (Cat No. 1217202) Elisa detection kits were purchased from DaKeWei. Bay 87-2243 (Cat No. S7309) was purchased from Selleck. Anti-NF-κB (Cat No. ab16502), anti-JNK (Cat No. ab179461), anti-Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cat No. ab17942), anti-CD31 (Cat No. ab134168), anti-Ki67 (Cat No. ab15580), anti-PCNA (Cat No. ab92552), and Anti-beta Actin (Cat No. ab8226) antibodies were purchased from Abcam. Anti-p-STAT3 (Tyr705) (Cat No. 9145), anti-STAT3 (Cat No. 9139) and anti-F4/80 (Cat No. 70076) were purchased from Cell Signaling Technology.
8
Culturing Human Umbilical Vein Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human umbilical vein endothelial cells (HUVECs; ScienCell, Carlsbad, CA, USA) were cultured in endothelial cell media (ScienCell, Carlsbad, CA, USA) with 20% FBS and 1% of endothelial cell growth supplement (ECGS), 1% penicillin (100 μg/mL), and streptomycin (100 μg/mL) and incubated at 37 °C in a 5% CO2 atmosphere. HUVECs were passaged when the cells reached 90% confluence and used for subsequent experiments at passages 4–7.
9
Irradiation of Human Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Human artery endothelial cell line (HAEC) was procured from iCell Bioscience Inc. (iCell-0015a) and maintained in endothelial cell media (ScienCell). The culture medium was supplemented with 5% fetal bovine serum (FBS), 1% growth factors, and 1% penicillin-streptomycin solution. Culturing was carried out at 37°C in an environment with 5% humidity. The EA.hy926 cell line, acquired from the Cell Bank of Type Culture Collection of the Chinese Academy of Sciences, was cultured in Dulbecco's modified Eagle's medium (DMEM, Hyclone, USA) supplemented with 1% penicillin-streptomycin solution (Hyclone, USA) and 10% FBS (Gibco, USA). This cell line was also maintained at 37°C in an atmosphere with 5% humidity. Both cell lines were utilized for experimentation within passages 5–15. Irradiation of the cells was carried out using a KUBTEC XCELL 225 X-ray cabinet. According to the multiple low-dose irradiation to animals, the radiation dose for the injury effect of abdominal aortic endothelial cells was determined to be 187.5, 375, and 750 mGy, delivered at a dose rate of 108 mGy/min.
10
Endothelial and Fibroblast Cell Proliferation Assay
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!