The HL-1 rat cardiomyocytes were cultured on the NWMEA biodevice to test the cellular electroporation and record electrical potential. In the preparation before cell culture, the NWMEA biodevice was incubated overnight with 5 µg/mL fibronectin in 0.02% gelatin solution at 4 °C to enhance cell attachment on the device surface. During the cell culture, the frozen HL-1 cardiac muscle cell line (Louisiana State University Health Science Center, USA) was thawed in a 37 °C water bath and incubated in an HL-1 expansion medium to recover the cardiomyocytes. To purify the HL-1 cardiomyocytes, they were pelleted by centrifuging the HL-1 expansion medium including the cells at 300 × g for 2–3 min and resuspended into 10 to 15 mL of fresh HL-1 expansion medium. The HL-1 cardiomyocyte suspension was transferred into the NWMEA biodevice at a density of 104 cells/cm2 and maintained in a standard incubator at 37 °C and 5% CO2 for the subsequent experiments. The HL-1 expansion medium was made from a mixture of 43.5 mL Claycomb basal medium (51800 C, Sigma-Aldrich), 5 mL qualified FBS (TMS-016-B, EMD Millipore), 0.5 mL l -glutamine at 200 mM (A0937, Sigma-Aldrich), 0.5 mL norepinephrine at 10 mM (TMS-002-C, EMD Millipore) and 0.5 mL penicillin/streptomycin at 100X (TMS-AB2-C, EMD Millipore). The cardiomyocyte culture was imaged by an upright widefield microscope (Olympus BX63, Japan).
Tms ab2 c
Manufactured by Merck Group
Sourced in United States, Germany
The TMS-AB2-C is a laboratory equipment product manufactured by Merck Group. It is designed to perform a core function within a laboratory setting. However, a detailed description of the product's features and capabilities cannot be provided in an unbiased and factual manner while maintaining conciseness. Therefore, the description for this product is not available.
Automatically generated - may contain errors
Lab products found in correlation
Tms 002 c, by Merck Group (7 mentions)
Tms 001 c, by Merck Group (4 mentions)
Es 009 b, by Merck Group (3 mentions)
Dmem f12, by Merck Group (2 mentions)
Esg1107, by Merck Group (2 mentions)
Freezone 2.5 liter freeze dry system, by Labconco (2 mentions)
Es 009 c, by Merck Group (2 mentions)
Slm 220 b, by Merck Group (2 mentions)
Lif2010, by Merck Group (2 mentions)
Tms 016 b, by Merck Group (1 mentions)
A0937, by Merck Group (1 mentions)
Dmem nutrient mixture f 12, by Merck Group (1 mentions)
Dispase 2 enzyme, by Merck Group (1 mentions)
Collagenase a, by Merck Group (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Penn strep, by Thermo Fisher Scientific (1 mentions)
Sh30071, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Accutase, by Merck Group (1 mentions)
12 protocols using tms ab2 c
1
Culturing HL-1 Cardiomyocytes on NWMEA Biodevice
2
Investigating the Effects of S1 Protein on Cardiomyocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human primary ventricular cardiomyocytes (AC16; Millipore, SCC109) were cultured in Dulbecco’s Modified Eagle Medium (DMEM)/Nutrient Mixture F-12 (Sigma, D6434) containing 2 mM L-glutamine (catalog no. TMS-002-C; EMD Millipore), 12.5% fetal bovine serum (catalog no. ES-009-B; EMD Millipore), and 1X penicillin–streptomycin solution (catalog no. TMS-AB2-C; EMD Millipore). AC16 cells (passages 7–9) were treated with two concentrations (0.1 and 1 nM) of S1 (Catalog no. Z03485; Genscript biotech, Piscataway, NJ, USA) with or without anti-ACE2 neutralizing antibody (Catalog no. 10108-MM37, Sino Biological Hong Kong, People’s Republic of China), anti-CD147 neutralizing antibody, and Mitotempo (GenScript Biotech, NJ, USA) for 24, 48, and 72 h (h). Untreated cells were used as the control.
3
Isolation of Mouse Corneal Stromal Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The removed mouse eyeballs were washed with sterile PBS containing penicillin and streptomycin (100 U/mL, TMS-AB2-C; Merck Millipore, Boston, MA, USA) three times. Each eyeball was digested for 12 hours at 4°C with 150 µL dispaseII enzyme (15 mg/mL; Sigma-Aldrich). Next, the corneal tissues were cut into small fragments after the removal of loose corneal epithelium. Furthermore, the tissues were digested with collagenase A (10 mg/mL; Sigma-Aldrich) at 37°C for 1 hour. Then, corneal stromal cells were suspended in a 25-cm2 culture flask with DF12 medium (DMEM F-12; Sigma-Aldrich). Adherent cells were observed after 10 to 12 hours at 37°C.
4
Silencing Human Per2 in AC16 Cardiomyocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The AC16 human cardiomyocyte cell line was purchased from the Millipore Sigma and cultured in 100 mm cell culture dishes with DMEM/F12 (Sigma, D6434) containing 12.5% FBS (EMD Millipore, ES-009-B), 2 mM l -Glutamine (EMD Millipore, TMS-002-C), and 1X Penicillin–Streptomycin Solution (EMD Millipore, TMS-AB2-C) at 37 °C, 5% CO2 and 90% humidity, based on the manufacturer's protocols. AC16 cells were plated in 12-well plate at 1.0X105 cells/well or in 96-well plate at 1.0X104 cells/well. Twenty-four hour later, cells were transfected with human Per2 or control siRNAs (Life Technologies) using Lipofectamine 2000 (Life Technologies). After one day of transfection, normal AC16 growth medium was added. The cells were allowed to incubate for an additional day and then used for experiments. Different doses of Per2 siRNA (10, 20, and 30 pmol) were used to determine their effects on transcript levels of Per2 and the dose of 20 pmol was selected for subsequent trials in this study.
All cell culture studies were performed in accordance with our approved protocol by the Institutional Biosafety Committee of Indiana University.
All cell culture studies were performed in accordance with our approved protocol by the Institutional Biosafety Committee of Indiana University.
5
Culturing Mouse and Drosophila Cell Lines
6
Culturing Yeast, Caulobacter, and Mouse ESCs
7
Culturing Human Fetal Osteoblasts and Embryonic Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A human fetal osteoblast cell line obtained from the Cell Applications Inc. (406K-05f) was used. Cells were maintained in McCoy's 5A medium (Gibco, 16600082) which was supplemented with heat-inactivated 10% FBS (Invitrogen, 10108-157), 1% Antibiotic-Antimycotic (Gibco, 15240062), 50 mg/ml L-Ascorbic Acid Phosphate and Magnesium Salt (Fujifilm Wako Chemical corporation, 013-19641). As a comparison for immuno-staining, clinical grade human embryonic stem cell (hESC) line KCL-033, a kind gift from Dr Tamir Rashid (King's College London), was used. hESCs were deemed a good comparison because of its low actin and vinculin expression on compliant substrates22 (link),24 (link),25 (link). hESCs were cultured on Geltrex and maintained in mTESR Plus media (Stemcell Technologies, 05825) supplemented with 1% penicillin and streptomycin (Sigma, TMS-AB2-C). Accutase (Sigma, A6964) was used to detach both the fHObs and hESCs for passaging and seeding. Passage 3–5 fHObs and passage 25–30 hESCs were used for all experiments. Seeding densities were 40,000 cells per well in a 96 well plate or 400,000 cells per well in a 4 well plate.
8
Thermal Gelation of Human Serum Albumin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human albumin (Sigma A1653) was dissolved in either deionized water, DBPS (Sigma, D8537), or DMEM/F12 (Fisher Scientific, 11574546). DMEM/F12 was supplemented with 1% Penicillin/Streptomycin (Sigma TMS-AB2-C). Human albumin dissolved in DMEM/F12 pre-warmed to 37 °C to make up a 10% (w/v) human albumin solution was used for in vitro experiments. Once filtered, a small volume from a sterile stock solution of 5 M NaCl (Sigma, S5886) was added to the 10% HSA to raise the molarity of NaCl by 30 mM, giving a final approximate NaCl molarity of 150 mM. The solution was then syringe filtered with a 0.1 µm syringe filter (Millipore SLVV033RS) for sterility.
Depending on the experiments, the final solution was added to either 96 well plates (Falcon 351172, 50 µl solution added) or 4 well plates (Thermoscientific 179830, 500 µl solution added) and sealed with aluminium plate sealers (Greiner Bio-One, 676090). Samples were then transferred into an oven and the temperature was raised to 73 °C. Samples were left for 4 h to gel and at the end of the heating process, translucent HSA hydrogels were obtained.
Depending on the experiments, the final solution was added to either 96 well plates (Falcon 351172, 50 µl solution added) or 4 well plates (Thermoscientific 179830, 500 µl solution added) and sealed with aluminium plate sealers (Greiner Bio-One, 676090). Samples were then transferred into an oven and the temperature was raised to 73 °C. Samples were left for 4 h to gel and at the end of the heating process, translucent HSA hydrogels were obtained.
9
Cell Culture Reagents and Techniques
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following reagents were used: α-MEM (Sigma, M2279), 0.25% Trypsin-EDTA (Sigma, T3924), Fetal Bovine Serum (Millipore, ES-009-B), L-Glutamine (Millipore, TMS-002-C), Penicillin-Streptomycin solution (Millipore, TMS-AB2-C), Hygromycin B (Sigma, H0654-500MG) and miRVana kit (Ambion, AM1560).
10
Cell Line Cultivation and Maintenance
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell lines including Huh7, HepG2, and 293T were obtained from the Shanghai Cell Bank of the Chinese Academy of Sciences and stored in liquid nitrogen in our laboratory. These cells were cultured in DMEM (C11995500BT, Gibco, USA) medium supplemented with 10% fetal bovine serum (10099-141, Gibco, USA) and 1% penicillin/streptomycin (TMS-AB2-C, Sigma, Germany). They all grew in good condition with normal morphology.
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!