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Live dead cytotoxicity assay for mammalian cells

Manufactured by Thermo Fisher Scientific
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The LIVE/DEAD cytotoxicity assay for mammalian cells is a fluorescence-based kit used to determine the viability of cells. The assay uses two fluorescent dyes: one that stains live cells and another that stains dead cells. The kit provides a simple, rapid, and quantitative method to assess the cytotoxicity of various treatments or conditions on mammalian cell cultures.

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3 protocols using live dead cytotoxicity assay for mammalian cells

1

Cytotoxicity Evaluation of Hydrogels

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Cytotoxicity test was performed, as has been mentioned elsewhere by Thi et al. [31 ], towards HDF via LIVE/DEAD cytotoxicity assay for mammalian cells (Thermo Fisher Scientific, Waltham, MA, USA). The hydrogels (n = 3) were fabricated in a 48-well polystyrene culture plate by using sterile gelatin and genipin solution. Immediately after polymerisation, 5 × 104 HDF passage three were seeded on the top of hydrogel prior to the incubation for 24 h. Cell toxicity was examined by using a fluorescence microscope (Nikon A1R-A1, Japan) at 100× magnification after treatment with 500 µL of a mixture of 2 mM acetomethoxy derivate of calcein (calcein-AM) and 4mM ethidium homodimer-1 (EthD-1) at 37 °C for 30 min.
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2

Investigating Photosensitizer Effects on Cells

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Fmoc-Phe-Phe peptide powder with a purity greater than 95% was purchased from Bachem (Bubendorf, Switzerland). The porphyrins H2-T(MePy)P(I4), Zn-T(MePy)P(I4), and Zn-T(MePy)P(Cl4) were synthesized as previously described [43 (link)]. The NIH3T3 cell line was cultured at 37 °C, 5% CO2 in DMEM (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) medium supplemented with 10% fetal bovine serum (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) and 50 μg/mL gentamycin (Applichem, Darmstadt, Germany). Thiazolyl blue tetrazolium bromide (MTT), isopropanol, dimethylsulfoxide (DMSO), ethanol, hexamethyldisilane (HMDS), and phosphate-buffered saline (PBS) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Live/Dead cytotoxicity assay for mammalian cells was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Tissue culture (TC) inserts with a 0.4 μm pore size were purchased from Sarstedt (Nümbrecht, Germany).
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3

3D Bioprinted Hydrogel Viability Assay

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The encapsulated cells in the hydrogels were tested for cell viability using a Live/Dead Cytotoxicity assay for mammalian cells (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s recommendations. Sterilized bioinks were prepared by fabricating the sterilized GE, PVA, and GNP powder and were printed using a 3D bioprinter. The bioinks were encapsulated with HDFs with 1.5 × 106 per mL of cell density. Cell viability will be assessed 24 h after post-printing. After 30 min of treatment with 250 μL of a mixture of 2 mM acetoxymethoxy calcein derivate (calcein-AM) and 4 mM ethidium homodimer-1 (EthD-1) at 37°C, cell toxicity was assessed using a fluorescence microscope (Nikon A1R-A1, Japan) at ×100 magnification.
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