A 3D cell culture model of human keratinocytes was purchased from MatTek Corporation (Ashland, MA). Irritation testing was performed according to manufacturer’s protocol. Briefly, upon arrival of the kit, fresh media was replaced and tissue inserts were incubated overnight at 37 °C with 5% CO2. The next day, tissues were dosed with 30 μL of saline (negative control), 30 μL of 5% sodium dodecyl sulfate (positive control), 25 mg of PTTC, and 30 μL of PTTC emulsion (n = 3 for each group). After incubating for one hour, the surface of the tissues was washed thoroughly with saline solution to remove any residual solution. The tissue inserts were incubated again for approximately 24 h. MTT reagent was added and allowed to incubate for 3 h. Absorbance was measured at 340 nm. Cell viability was calculated using a spreadsheet provided by MatTek; viability less than 50% was determined to be irritant.
3d cell culture model of human keratinocytes
Manufactured by MatTek
The 3D cell culture model of human keratinocytes is a laboratory equipment used for the in vitro culture of human skin cells. It provides a three-dimensional environment that mimics the natural structure and function of human epidermis, allowing for the study of cell-cell and cell-matrix interactions.
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2 protocols using 3d cell culture model of human keratinocytes
1
Irritation Testing of a 3D Keratinocyte Model
2
Keratinocyte 3D Culture Irritation Assay
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A 3D cell culture model of human keratinocytes was purchased from MatTek Corporation. Irritation testing was performed according to manufacturer’s protocol. Briefly, upon arrival of the kit, fresh media was replaced and tissue inserts were incubated overnight at 37 °C with 5% CO2. The next day, tissues were dosed with 30 µL of saline (negative control), 30 µL of 5% sodium dodecyl sulfate (positive control), and 30 µL of glycopyrrolate solution (n = 3 for each group). After incubating for 1 h, the surface of the tissues was washed thoroughly with saline solution to remove any residual solution. The tissue inserts were incubated again for approximately 24 h. MTT reagent was added and allowed to incubate for 3 h followed by isopropanol extraction for 2 h. Absorbance was measured at 340 nm. Cell viability was calculated using a spreadsheet provided by MatTek; viability less than 50% was determined to be irritant.
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