Primary neonatal rat cardiomyocytes (NRCMs) were obtained from 1- to 2-day-old Sprague-Dawley rats [18 ]. Removed and digested the heart in PBS containing collagenase type II (Worthington, NJ, USA) and pancreatin (Sigma, MO, USA). Deserted the atria and great vessels. Minced and digested the ventricles with collagenase type II and pancreatin. Collected and cultured Cells from digestion in DMEM (GIBCO, Invitrogen Inc.) for 2–4 h to decrease fibroblasts and increase cardiomyocytes. Cultured the cardiomyocytes at 37 °C with 5% CO2. The cells were received treatment when the cells were grown to 80–90% confluence. Firstly, divided and treated the primary cardiomyocytes with PBS, Ang II (10−6 M; Sigma), tumstatin (69–88) (10−6 M) and Ang II + tumstatin (69–88), respectively. Secondly, the primary cardiomyocytes were divided and treated with PBS, Ang II, Ang II + tumstatin (69–88), diethyldithiocarbamate (DETC; 10−6 M; Sigma) and DETC + Ang II + tumstatin (69–88), respectively. Thirdly, divided and treated the primary cardiomyocytes with PBS, Ang II, Tiron (antioxidant; 10−5 M; Sigma) and Ang II + Tiron. All the reagents added into the medium ate the same time for 24 h.
Tumstatin
Manufactured by Merck Group
Sourced in United States
Tumstatin is a laboratory equipment product manufactured by Merck Group. It is a type IV collagen-derived protein that functions as a potent inhibitor of angiogenesis, the formation of new blood vessels. The core function of Tumstatin is to enable researchers to study the mechanisms and applications of this anti-angiogenic property in various scientific and medical research contexts.
Automatically generated - may contain errors
Lab products found in correlation
Ang 2, by Merck Group (1 mentions)
Tiron, by Merck Group (1 mentions)
Pancreatin, by Merck Group (1 mentions)
Collagenase type 2, by Worthington (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s minimum essential medium dmem, by Thermo Fisher Scientific (1 mentions)
Tnf α, by Merck Group (1 mentions)
Saos 2 osteosarcoma cell line, by American Type Culture Collection (1 mentions)
2 protocols using tumstatin
1
Primary Rat Cardiomyocytes Treatment
2
Tumstatin and TNF-α Regulation of Osteosarcoma
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Tumstatin and TNF-α were purchased from Sigma-Aldrich (St. Louis, MO, USA). The stock solution of Tumstatin was prepared in dimethyl sulfoxide and stores at -15˚C before use in the experiment. Fetal bovine serum (FBS) and Dulbecco's modified Eagle's minimum essential medium (D-MEM) were obtained from Gibco-BRL (Gaithersburg, MD, USA).
Cell culture. Saos-2 osteosarcoma cell line and human embryonic kidney HEK293 cells were obtained from the American Type Culture Collection (ATCC; Rockville, MD, USA). The cells were cultured in D-MEM containing 10% FBS, 2 mM glutamine, 100 U/ml penicillin and 100 µg/ml streptomycin.
Cell culture. Saos-2 osteosarcoma cell line and human embryonic kidney HEK293 cells were obtained from the American Type Culture Collection (ATCC; Rockville, MD, USA). The cells were cultured in D-MEM containing 10% FBS, 2 mM glutamine, 100 U/ml penicillin and 100 µg/ml streptomycin.
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