The DRG tissues were dissected and incubated with D-Hanks solution (Beyotime Institute of Biotechnology, Shanghai, China). The mixture was digested with 0.25% trypsin (Beyotime Institute of Biotechnology) at 37°C for 20 min. A pipette was used to mix 20 ml Dulbecco’s modified Eagle’s medium with Ham’s F-12 (DMEM/F12; Gibco; Thermo Fisher Scientific, Inc.) in a proportion of 1:1, and fetal bovine serum (FBS; Sigma-Aldrich; Merck KGaA) was then added into the mixture to stop the digestion process. Then, the cell suspension was collected. The suspension was centrifuged at 224 × g for 15 min, and the supernatant was discarded. A total of 10 ml DMEM/F12 was added into the precipitate. The cell debris and residual tissues were sunk, and the cell suspension was collected. Cells were seeded into a polylysine pre-coated culture flask (75 cm2, 250 ml) and placed in an incubator with 5% CO2 at 37°C. After 48 h, the cells that attached on the pre-coated flask were rat DRG neurons.
D hanks solution
Manufactured by Beyotime
Sourced in China
D-Hanks solution is a sterile, cell culture medium designed for the maintenance and growth of various cell lines. It contains a balanced salt solution, amino acids, vitamins, and other essential components to support cell survival and proliferation in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Trypsin, by Beyotime (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Bca protein concentration determination kit, by Solarbio (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions)
Anti β catenin, by Abcam (1 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
Collagenase type 2, by Merck Group (1 mentions)
Irdye 680cw, by LI COR (1 mentions)
5 bromo 2 deoxyuridine 5 brdu, by Merck Group (1 mentions)
3 protocols using d hanks solution
1
Rat DRG Neuron Isolation Protocol
2
Modeling Ischemia-Reperfusion Injury in H9C2 Cells
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The D-galactose induction was conducted as previously described (18 (link)). D-galactose (10 g/l) was added to H9C2 cells for 48 h at 37°C. Hypoxic conditions were produced using D-Hank's solution (Beyotime Institute of Biotechnology) saturated with 95% N2 and 5% CO2 at 37°C. The pH was regulated to 6.8 using lactate to mimic ischemic conditions. Aged H9C2 cells were put into a hypoxic incubator which was equilibrated with 1% O2, 5% CO2 and 94% N2 at 37°C. Following the hypoxia period, the culture medium was rapidly replaced with fresh DMEM with 10% FBS (normoxic culture solution) to mimic reoxygenation.
3
Isolation and Culture of Chondrocytes
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The suppliers and the catalog numbers of the reagents are as follows. Dulbecco’s modified Eagle’s medium (DMEM) (Life Technologies, Carlsbad, CA, United States ; 22,400–089), fetal bovine serum (Life Technologies; 10,099), antibiotic-antimycotic (Life Technologies; 15,240–112), phosphate-buffered saline (PBS) (Life Technologies; 10010-049, pH7.4), trypsin-EDTA (Life Technologies; 25300-054, 0.05%), bovine serum albumin (Life Technologies; 15560012); D-Hanks solution (Beyotime, Jiangsu, China; C0218); type II collagenase (Sigma, St. Louis, MO, United States ; C6885); 5-bromo-2'-deoxyuridine (5-BrdU) (Sigma; B5002); Cell Counting Kit-8 (CCK-8; DOJINDO, Kumamoto, Japan; CK04), IRDye 680CW (Licor, Lincoln, NE, United States ), protein molecular weight markers (Beyotime; P0066), BCA protein concentration determination kit (Solarbio, Beijing, China; PC0020). Antibodies: anti-glycogen synthase kinase beta (GSK3β) (Cell Signaling Technology (CST), Danvers, MA, United States ; 12,456), anti-AKT serine/threonine kinase 1 (AKT1) (CST; 2,967), anti-AKT serine/threonine kinase 2 (AKT2) (CST; 2,962), anti-β-catenin (Abcam, Cambridge, MA, United States ; ab24925), and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Abbkine, Wuhan, China; Abp57259).
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