The cells were cultured in mTeSR™1 (StemCell Technologies, Van-couver, BC, Canada) and coated with human vitronectin (Gibco, Gaithersburg, MD, USA). Cells were passaged by 1:10 with 0.02% EDTA (Sigma, Darmstadt, Germany). The 0.2 g EDTA was dissolved in 1 Liter Dulbecco’s phosphate-buffered saline (DPBS, Biological industries, Cromwell, CT, USA).
Dulbecco s phosphate buffered saline dpbs
Manufactured by Sartorius
Sourced in Israel
Dulbecco's phosphate-buffered saline (DPBS) is a balanced salt solution commonly used in biological research and cell culture applications. It is designed to maintain the pH and osmotic balance of cells in vitro. DPBS is a sterile, isotonic solution that provides a physiologically compatible environment for cells.
Automatically generated - may contain errors
Lab products found in correlation
Human vitronectin, by Thermo Fisher Scientific (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Sartorius (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
Mtesr1, by STEMCELL (1 mentions)
Biotinamidohexanoic acid n hydroxysuccinimide ester, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Trypsin edta solution, by Sartorius (1 mentions)
L glutamine, by Sartorius (1 mentions)
4 protocols using dulbecco s phosphate buffered saline dpbs
1
Culturing Cells in mTeSR™1 with Vitronectin
2
Biotinylation of Collagen Membranes
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Bio-Gide® collagen membranes (Geistlich Pharma, Wolhusen, Switzerland) were cut into 6 mm diameter discs using a disposable biopsy punch. All discs were labeled with 3 mg/mL biotinamidohexanoic acid N-hydroxysuccinimide ester (MERCK, Darmstadt, Germany) for 1 h at room temperature, washed three times with Dulbecco’s phosphate-buffered saline (DPBS, calcium/magnesium- free, pH 7.4, Biological Industries, Beit-Haemek, Israel) for 12 h and dried overnight.
3
Mesenchymal Stromal Cell Isolation and Characterization
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Mouse mesenchymal stromal cells (mMSC) were isolated, cultured and characterized as previously reported [14 (link)]. Cells were kindly provided by Dr. Osnat Almogi-Hazan, Hebrew University. Bovine mesenchymal stromal cells (bMSC), as we have previously reported [4 (link)], were isolated, cultured and characterized based on generally accepted criteria [3 (link), 15 (link)]. For some experiments and microscopic imaging, cells were preincubated with carboxyfluorescein succinimidyl ester (CFSE; Celltrace, Invitrogen). Briefly, culture medium was removed and cells were treated with 10 μM CFSE in Dulbecco’s Phosphate Buffered Saline (DPBS, Biological Industries, Beit Haemek, Israel) and incubated for 20 min at 37°, 5% CO2. Cells were washed 3 times with a culture medium containing 10% FBS (Gibco), and incubated for further 10 min before harvesting or further processing.
4
Cell Viability and Oxidative Stress Assays
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The following reagents were obtained from Sigma-Aldrich (St. Louis, MO, USA): 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), Dimethyl sulfoxide (DMSO), vitamin C, L-3,4-dihydroxyphenylalanine (L-DOPA), Triton X-100, nitro blue tetrazolium (NBT) riboflavin, ethylenediaminetetraacetic acid (EDTA), thiobarbituric acid (TBA), trichloroacetic acid (TCA), 2′,7′-dichlorofluorescin diacetate (DCFH-DA).
The Muse® Annexin V & Dead Cell Kit was obtained from Luminex (Luminex Corporate, Austin, TX, USA). Additionally, Dulbecco’s Modified Eagle’s Medium (DMEM), Fetal Bovine Serum (FBS), Penicillin-Streptomycin solution, Trypsin-EDTA solution, L-glutamine, and Dulbecco’s Phosphate Buffered Saline (DPBS) were obtained from Biological Industries, Kibbutz Beit Haemek, Israel. DNX was provided from a purified (Figure S1 ) stock produced by Seda KILINÇ in a joint venture between Erzincan Binali Yıldırım University and Marmara University [29 ].
The Muse® Annexin V & Dead Cell Kit was obtained from Luminex (Luminex Corporate, Austin, TX, USA). Additionally, Dulbecco’s Modified Eagle’s Medium (DMEM), Fetal Bovine Serum (FBS), Penicillin-Streptomycin solution, Trypsin-EDTA solution, L-glutamine, and Dulbecco’s Phosphate Buffered Saline (DPBS) were obtained from Biological Industries, Kibbutz Beit Haemek, Israel. DNX was provided from a purified (
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