At the Society of Toxicology 56th Annual Meeting and ToxExpo™, Trevigen Inc. described its progress towards the development of an in vitro Genetic Toxicology Skin Assay. Since our skin is exposed continuously to a cocktail of various environmental agents, including cosmetics, dermal care products, both personal and industrial cleaning agents, household products, fertilizers, pesticides, industrial chemicals and by–products, air pollution, etc; a high-throughput reliable assay to assess genotoxic risk to environmental exposure has long been a goal of the community. The technology is the subject of a Trevigen pending patent and was supported by NIEHS R41ES026908.
It has been shown that risk assessment data from the micronucleus assay together with the traditional comet analysis of cells isolated from treated 3D skin culture models is a better predictor of skin toxicity compared to the standard battery of in vitro toxicity assays. However, in some in vitro models, unacceptable backgrounds were observed as a function of skin preparation. This may be a result of the stage of differentiation of the skin and associated DNA damage when assays are performed. One would expect higher comet backgrounds in cells migrating toward the stratum corneum.
Trevigen researchers have combined extracellular matrix 3D Culture technology with the CometChip® technology, exclusively licensed from MIT, to develop a platform to specifically isolate basal keratinocytes for genetic toxicology testing using in vitro skin models. The elimination of cells migrating to the stratum corneum, that potentially harbor damaged DNA from the analysis, reduces the background for some 3D models.
Patent pending DermaChip® Technology
DermaChip® is a disposable gel attached to a specially treated glass slide, indexed for 96 well locations, containing thousands of 30 micron sized pores designed to capture and array cells. The gel specifically traps basal keratinocytes expressing integrin beta-1 before they differentiate and migrate to the stratum corneum. During their natural migration, cells stop dividing and ultimately lose their nucleus. During the process, DNA damage accumulates resulting in high comet backgrounds. By trapping basal keratinocytes, we can eliminate this background. Prior to comet analysis, the identity of trapped cells is confirmed by Anti-integrin beta-1 immunostaining labeled with quantum dots. Only basal keratinocytes are included in comet data analysis.
Trevigen will submit a Phase II SBIR application to continue the development of this technology. We plan to evaluate a library of compounds that have known genetic toxicology profiles and further develop our comet analysis software to include a DermaChip® specific module. Parties interested in collaborating with Trevigen on the project are encouraged to contact Jay George, Ph.D., Chief Scientific Officer at Trevigen Inc., firstname.lastname@example.org.
CometChip® can dramatically improve lab productivity. To find out more about the CometChip® Platform, visit: https://www.trevigen.com/cat/1/56/0/CometChip/