In this study, the bend.3 mouse brain microvascular endothelial cell line was purchased from Shanghai Zhongqiao Xinzhou Biotechnology Co., Ltd., and cultured in high glucose Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin. RIF was purchased from Sangon Biotech (Shanghai, China) Co., Ltd. The ANG-peptide (coupled with FITC) was purchased from the Chinese Peptide Company (Hangzhou, China). All other cell culture media and reagents were purchased from Gibco (Carlsbad, CA, USA). Cell culture plates and transwell chamber were obtain from NEST Biotechnology Co. Ltd (Wuxi, China).
Cell culture plates
Manufactured by NEST Biotechnology
Sourced in China
Cell culture plates are a fundamental tool used in biomedical research and cell biology laboratories. They provide a sterile, controlled environment for the growth and maintenance of cells, tissues, or other biological samples. These plates are typically made of plastic and come in a variety of sizes and well configurations to accommodate different experimental needs.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Centrifuge tubes, by NEST Biotechnology (2 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (2 mentions)
Transwell chamber, by NEST Biotechnology (1 mentions)
Endothelial cell growth supplement (ecgs), by ScienCell (1 mentions)
α minimum essential medium α mem, by Thermo Fisher Scientific (1 mentions)
P erk, by ABclonal (1 mentions)
P jnk, by ABclonal (1 mentions)
Trap staining kit, by Merck Group (1 mentions)
Sirt3, by ABclonal (1 mentions)
Cck 8, by Dojindo Laboratories (1 mentions)
Penicillin streptomycin, by Solarbio (1 mentions)
M csf, by R&D Systems (1 mentions)
Rankl, by R&D Systems (1 mentions)
Gapdh, by ABclonal (1 mentions)
Catalase, by Sangon (1 mentions)
Goat anti gfap, by Abcam (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Hoechst 33258, by Merck Group (1 mentions)
Glucose oxidase, by Merck Group (1 mentions)
8 protocols using cell culture plates
1
Bend.3 Mouse Brain Microvascular Endothelial Cell Culture
2
Endothelial Cell Dysfunction in Hyperglycemia
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Primary hRECs were purchased from PromoCell (C-12200, Heidelberg, Germany), and all of the experiments were performed using 2-5 passages of hRECs. hRECs were grown in complete endothelial culture medium ECM (ScienCell, 1001) containing 1% endothelial cell growth supplement (ScienCell, 1052), 5% fetal bovine serum (Gibco, A3160802), and 1% penicillin/streptomycin solution (Gibco, 15140-122) at 37°C in a humidified atmosphere of 5% carbon dioxide. Cell culture plates and centrifuge tubes were purchased from NEST Biotechnology Co. Ltd. (Wuxi, China). For high glucose cells, experiments indicated that the amount of D-glucose (MCE, HY-B0389) was added directly in ECM media to obtain a final concentration of 10, 15, 20, or 30 mM, and a hypertonic group (24.5 mM mannitol and 5.5 mM glucose) was added to exclude hyperosmolarity effects. To detect the role of ferroptotic signals in HG-induced endothelial dysfunction, hRECs were treated with 10 μM apoptosis inhibitor tauroursodeoxycholic acid (TUDCA) (MCE, 35807-85-3), 10 μM necrosis inhibitor necrostatin-1 (MCE, 4311-88-0), 10 μM ferroptosis inhibitor ferrostatin-1 (Fer-1) (MCE, HY-100579), and 10 μM pyroptosis inhibitor tetraethylthiuram disulfide (TETD) (MCE, 97-77-8) for 48 h.
3
Osteoclastogenesis Regulation Protocol
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VPA was obtained from MedChemExpress (New Jersey, USA). GAPDH, JNK, p-JNK, p38, p-p38, ERK, p-ERK, SIRT3, CD3, LY6G and CX3CR1 antibodies were purchased from ABclonal (Wuhan, China), and antibodies against CX3CL1 and Lamp-2b were bought from Abcam. Osteocalcin antibody was purchased from Takara (Japan, M173). Penicillin-streptomycin was purchased from Solarbio (Beijing, China). The RANKL and M-CSF were bought from R&D Systems (Minnesota, USA). The cell culture plates were from NEST (Jiangsu, China). Minimum Essential Medium Alpha (α-MEM) and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA). CCK-8 (Kumamoto, Japan) was bought from Dojindo. The enzyme-linked immunosorbent assay (ELISA) kits were purchased from CUSABIO and Elabscience (Wuhan, China). TRAP staining kit was obtained from Sigma–Aldrich (St. Louis, MO, USA).
4
Lentiviral Vector Production and Transduction
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Lentivirus plasmids, empty backbone of pLenti CMV Blast DEST (plasmid #17451) addgene, pLenti CMV Puro DEST (plasmid #17452) addgene, psPAX2, pMD2.G, 293 T cells, N2A cells, XL 1 E. Coli, DMEM (Gibco), FBS (BioExcel), Blastidine (Invivogen company), puromycin (Invivogen company), prestained marker (Yeaseen), streptomycin/penicillin solution (Gibco), anti-rabbit granzyme B (Abcam, cat. No. ab53097), anti-goat GFAP (Abcam, cat. No. ab53554), anti-hamster Serpina3n (Merck cat. No. MABC1182), anti-chicken MAP-2 (Abcam cat. No. ab5392), Hoechst 33258 (Sigma-Aldrich cat. No. 14530), anti-mouse 6his-tag (Proteintech, cat. No. 66005-1-Ig), anti-rabbit C-Myc (Proteintech, cat. No. 10828-1-AP), glucose oxidase (Sigma, cat. No. G7141), and catalase (Sangon, cat. No. A001847). Cell culture plates (Nest Biotechnology Co., Ltd.). Human granzyme B gene sequences were stored in our laboratory (EC:3.4.21.79).
5
Ultrapure Water-based Biochemical Assay
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Ultrapure water was produced by a Milli-Q system (Millipore, USA). HPLC-grade acetonitrile (ACN) (Fisher, USA) and trifluoroacetic acid (TFA) (Sigma, USA) were used. The BCA kit and western blot gel buffers were purchased from Beijing CellChip Biotechnology Company (Beijing, China). Cell culture plates were purchased from Nest Biotechnology Company (Jiangsu, China). All other reagents and chemicals (G418) were purchased from Sigma Chemicals Company (Sigma, USA).
6
Quantifying Neutrophil Adhesion with AGP
7
Isolation and Purification of sPLA2
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All chemical reagents used in the present study were analytical grades. Items purchased for isolation and purification of secretory phospholipase A2 are stated in a previous study (Abdullah et al., 2021 (link)). Culture medium and fetal bovine serum were purchased from Gibco® by Life Technologies™, Massachusetts, US. Accutase Cell Detachment Solution was purchased from Capricorn Scientific GmBH, Ebsdorfergrund, Germany. Retinoic acid was purchased from Sigma-Aldrich, Bornem, Belgium. Human BDNF was purchased from StemCell™ Technologies, British Columbia, Canada. Cell culture plates were purchased from NEST® Biotechnology Co., Ltd., Rahway, US.
8
Transfection of C918 Cells with S100A13 siRNA
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The adult retinal pigment epithelial cell line-19 (ARPE-19) and the human invasive UVM cell line (C918) were purchased from Procell Life Science&Technology Co., Ltd (Wuhan, Hubei, China). ARPE-19 cell was cultured in Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F12 Ham’s Liquid media (DMEM/F-12; Cytiva/Global Life Sciences Solutions, Marlborough, MA) containing 10% fetal bovine serum (FBS; Gibco, Carlsbad, CA, USA), along with 100 U/mL penicillin and streptomycin (Gibco, Carlsbad, CA, USA). C918 cell was cultured in Roswell Park Memorial Institute 1640 liquid media (RPMI1640; Gibco, Carlsbad, CA, USA) containing 10% fetal bovine serum (FBS; Gibco, Carlsbad, CA, USA), along with 100 U/mL penicillin and streptomycin (Gibco, Carlsbad, CA, USA). Cells were maintained in an incubator (Thermo Fisher Scientific, Waltham, MA) at 37°C, with 95% humidity, and 5% CO2. Cell culture plates, round coverslips, and centrifuge tubes were obtained from (NEST Biotechnology; Wuxi, China). C918 cells were transfected with the generated small interfering RNAs (RiboBio; Guangzhou, China) targeting gene S100A13 and its control siRNAs, according to the manufacturer’s procedure. The siRNA sequences for the gene S100A13 were ACTCGGAGCTCAAGTTCAA.
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