Using Lab-Grown Skin as an In Vitro Model

Objective

To compare sensitizing and non-sensitizing materials using an IL-18 / MTT based in vitro sensitization assay to determine Labskin’s response as an in vitro skin model.

Methods

  • Labskin Maintenance Medium (4.5 mL) was added to each well. The 12 well plate configuration was used to conduct the experiment.
  • Known sensitizing materials (4-Nitrobenzyl bromide [NBB] and Eugenol) along with known non-sensitizing material (Lactic Acid) were used to dose tissues (52 μL).
  • A 12 well plate was used to incubate tissues with 1 mL of MTT (1 mg/mL) for 3.5 hours.
  • In each 12 well plate insert, Isopropanol (2 mL) was placed and a further 2 mL Isopropanol was added to the top of the tissues. Extraction occurred overnight.
  • The next day spectrophotometer (OD 540-690) was used to read 200 μL aliquots for viability once removed. ELISA (MBL) was used to analyze the media for IL-18 secretion into the culture media.

Results

As expected, there was no increase in IL-18 with the non-sensitizer Lactic Acid in comparison to vehicle control (AOO [Acetone: Olive OIL]).

However, where a response should have been seen, there was no increase in IL-18 for the sensitizing material Eugenol in comparison to the vehicle control. Fold increases in IL-18 for NBB was detected.

IL-18 Secretion detected in culture media

Figure 1. IL-18 Secretion detected in culture media

Summary

In conclusion, this experiment demonstrated that Labskin full thickness in vitro skin model is able to respond to sensitizing/allergic materials.

About Labskin

At Labskin we deliver human skin microbiology services to support your product R&D activities in the cosmetic, personal care, medical device and pharmaceutical sectors. With our sector experience and use of technology, you will be accessing industry-focused services supported by world-leading skin science expertise.

Whether you need rapid, focused analysis or flexible, tailor-made research programs we can help you develop and validate skincare ingredients and products which really work.

Our skin model is a 3D human skin equivalent that incorporates vital biological components to model normal skin function.

Developed over 12 years with more than 30 scientific journal publications, it is made from young keratinocytes (human skin cells) and adult fibroblasts (metabolically-active, collagen-producing human skin cells).

“An ideal platform for basic or applied skin research, testing compounds or formulated products for the cosmetic, pharmaceutical and chemical sectors.”


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Last updated: Oct 28, 2019 at 4:42 AM

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