The cytotoxicity was studied using Live/Dead staining, AlamarBlue and PicoGreen assays. To reveal the contact cytotoxicity, the samples were seeded with MSC (1.5 × 105 cells per construct) and stained in 24 h with Live/Dead staining kit (04511, Sigma-Aldrich, USA) and Hoechst 33258 (94403, Sigma-Aldrich, USA). The prepared mounts were analyzed using a laser scanning confocal microscope LSM 880 equipped with an AiryScan module and GaAsP detector (Carl Zeiss, Germany). The elution test was carried out using AlamarBlue (DAL1025, Invitrogen, Thermo Fisher Scientific, USA) and PicoGreen assays (P11495, Invitrogen, Thermo Fisher Scientific, USA) on a plate spectrofluorimeter Victor Nivo (PerkinElmer, USA) as described elsewhere [16 , 17 (link)]. The extraction was performed using DMEM/F12 medium containing 5% FCSIII (HyClone, USA) and 1% penicillin–streptomycin (15140122, Gibco, Thermo Fisher Scientific, USA) by incubating in a CO2 incubator at a temperature of 37 °C for 24 h. Sodium dodecyl sulfate (SDS) (L3771, Sigma-Aldrich, USA) was used as a positive control.
Live dead staining kit
Manufactured by Merck Group
Sourced in United States
The Live/Dead staining kit is a laboratory tool used to distinguish between viable and non-viable cells in a sample. It contains fluorescent dyes that selectively stain live and dead cells, allowing for their identification and enumeration under a microscope or flow cytometer. The kit provides a simple and reliable method for assessing cell viability across various cell types and applications.
Automatically generated - may contain errors
Lab products found in correlation
Alamar blue, by Thermo Fisher Scientific (1 mentions)
Picogreen, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Gaasp detector, by Zeiss (1 mentions)
Hoechst 33258, by Merck Group (1 mentions)
Victor nivo, by PerkinElmer (1 mentions)
Airyscan module, by Zeiss (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Chloroform, by Avantor (1 mentions)
Dichloromethane dcm, by Avantor (1 mentions)
Trifluoroacetic acid tfa, by Thermo Fisher Scientific (1 mentions)
Toluene, by Tedia (1 mentions)
Trifluoroacetic acid d tfa d, by Thermo Fisher Scientific (1 mentions)
Tetrahydrofuran thf, by Tedia (1 mentions)
Alamarblue cell viability reagent, by Thermo Fisher Scientific (1 mentions)
Avicel ph 101, by Merck Group (1 mentions)
Lysozyme, by Merck Group (1 mentions)
Gamma globulin, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
5 protocols using live dead staining kit
1
Cytotoxicity Assessment of Biomaterials
2
Synthesis and Characterization of Poloxamer-Based Biomaterials
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
O,O’-Bis(2-aminopropyl) polypropylene glycol-block-polyethylene glycol-block-polypropylene glycol (Poloxamer, PLX, PPG–PEG–PPG, Mn = 900), l -alanine, hydrogen bromide (HBr/acetic acid), and a LIVE/DEAD staining kit were purchased from Sigma-Aldrich (St. Louis, MO, USA). Tacrolimus (FK506) was obtained from LC Labs (Woburn, MA, USA). Trifluoroacetic acid-d (TFA-d) and trifluoroacetic acid (TFA) were purchased from Alfa Aesar (Wall Hill, MA, USA). l -lysine-(Z) was commercially obtained from ACROS (Morris Plains, NJ, USA). Diethyl ether, hexane, and ethanol (95%) were purchased from Echo Chemicals (Toufen, Miaoli, Taiwan). Dimethyl sulfoxide (DMSO) was obtained from J.T. Baker. Dulbecco’s modified eagle’s medium (DMEM), fetal bovine serum (FBS), and antimycotics–antibiotics were purchased from Gibco (Grand Island, NY, USA). Toluene and tetrahydrofuran (THF) were obtained from TEDIA (Fairfield, OH, USA), and chloroform, dichloromethane (DCM), and N,N-dimethylformaide (DMF) were purchased from AVANTOR (Center Valley, PA, USA). All solvents were dried over CaH2 before used.
3
Cell Viability Assay of Microcrystalline Cellulose
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Microcrystalline cellulose, Avicel PH 101, (MCC), 2,2,6,6-Tetramethylpiperidine-1oxyl radical (TEMPO), NaBr, NaCl, NaClO, NaOH, PVA (average Mw 146,000–186,000 and degree of saponification 99.9%), dimethyl sulfoxide (DMSO), aqueous hydroxyl amine (50%), sodium dodecyl sulfate (SDS), ethanol, hydrochloric acid, acetic acid, iso-propanol, Dulbecco’s phosphate buffered saline (PBS), lysozyme, gamma-globulin, bovine serum albumin, and a live-dead staining kit were purchased from Sigma Aldrich (Stockholm, Sweden). A micro bicinchoninic acid (BCA) protein assay kit was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Keratinocyte serum free medium (KSFM) and its supplements were purchased from Life Technologies (Bleiswijk, the Netherlands). Alamar blue (AB) cell viability reagent was purchased from Thermo Fisher Scientific (Waltham, MA, USA). All the chemicals used were of reagent or analytical grade.
4
Gellan Gum-based Biomaterial Synthesis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Gellan Gum (GelzanTM CM) of low acyl degree, sodium hydroxide (NaOH), ethylendiamine (EDA), tetrabuthylammonium hydroxide (TBA-OH), bis(4-nitrophenyl)carbonate (4-NPBC), acetone, dimethylsulfoxide anhydrous (DMSOa), Dowex® 50WX8 hydrogen form, O-[2-(6-Oxocaproylamino)ethyl]-O′-methylpolyethylene glycol 2000 (PEG-aldehyde 2000), Live/dead staining kit, Dulbecco’s phosphate buffered saline (DPBS), and Dulbecco’s Minimum Essential Medium (DMEM) deuterium oxide (D2O) were purchased from Sigma-Aldrich (Milan, Italy). Hydrochloric acid (HCl) was purchased from Fluka (Milan, Italy).
CellTiter 96® AQueous One Solution Cell Proliferation Assay (MTS) was purchased from Promega (Milan, Italy). HCT-116 (human colon tumor) and MC3T3-E1 (murine preostoblastic cells) were purchased from Euroclone (Milan, Italy).
CellTiter 96® AQueous One Solution Cell Proliferation Assay (MTS) was purchased from Promega (Milan, Italy). HCT-116 (human colon tumor) and MC3T3-E1 (murine preostoblastic cells) were purchased from Euroclone (Milan, Italy).
5
Cell Culture Preparation for DEP Experiments
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human K562 cells were cultured in RPMI-1640 media at 37 °C, supplemented with 10% heat-inactivated FBS (Biosera, UK), 1% l -glutamine and 1% penicillin–streptomycin. Cells were washed in low conductivity DEP buffer medium containing 8.5% sucrose and 0.3% dextrose and resuspended in CPM at a final concentration of 106 cells/ml. HeLa cells were cultured in MEM with Earle’s salts and non-essential amino acids (Biosera, UK) supplemented with 10% heat-inactivated FBS (Sigma-Aldrich, UK), 1% l -glutamine and 1% penicillin–streptomycin (Sigma Aldrich, UK). At about 80% confluency, cells were trypsinized and washed twice in DEP buffer medium and resuspended in CPM at a final concentration of 106 cells/ml. HL-1 cells were cultured in Claycomb media supplemented with 10% heat-inactivated FBS, 1% penicillin–streptomycin, 1% l -glutamine and Norepinephrine (0.1 mM) (Biosera, UK). The cells were trypsinised at 100% confluence and washed twice in DEP buffer medium before resuspending in CPM at a final concentration of 106 cells/ml. Viability of cell suspensions was confirmed on control samples prior to experiment using a LIVE/DEAD Staining Kit (Sigma Aldrich, UK). Further viability was monitored with trypan blue (Sigma Aldrich, UK).
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!