Jackson, G., Kubilus, J., Klausner, M., Sheasgreen, J., Hayden, P. MatTek Corporation, Ashland, MA.

This study by scientists at MatTek Corp. demonstrated that MatTek’s EpiDerm in vitro 3-D human skin tissue equivalent and MatTek’s EpiOcular in vitro 3-D human corneal tissue equivalent can be produced in a 96-well high-throughput microwell plate format in a highly reproducible manner, enabling the use of this format to help eliminate sample throughput bottlenecks encountered in safety and efficacy testing of consumer products and topical therapeutics. Prior to introduction of new consumer products and cosmetics, or human testing of new therapeutics, animal experiments are traditionally utilized to screen for safety and efficacy. However, large numbers of lead candidates generated by modern high throughput technologies renders animal testing expensive and impractical. A growing need exists for high throughput in vitro models which can provide rapid, reliable safety and efficacy screening. This poster describes development of in vitro models of human epidermal and ocular cultures in 96-well high-throughput screening (HTS) formats compatible with robotic manipulation. The HTS models are derived from normal human cells cultured at the air/liquid interface in 96-well microporous membrane plates to produce three-dimensional organotypic cultures. Culture histology of both models was evaluated by H&E staining of formalin fixed paraffin sections. The epidermal model displays a stratified and cornified differentiated structure, similar to native epidermis. The 96-well HTS ocular cultures are stratified squamous epithelium typical of in vivo ocular epithelium. Well-to-well (intraplate) and plate-to-plate (interlot) variability, as determined by the MTT viability assay, was generally found to be less than 10%. The HTS ocular model was also evaluated for compatibility with the EpiOcular Prediction Equation developed for predicting in vivo Draize ocular irritation scores with MatTek Corporation’s standard EpiOcular product (OCL-200). When tested with various concentrations of chemicals including sodium dodecyl sulfate, Triton X-100, benzalkonium chloride, sodium hydroxide and others, the HTS ocular model gave predicted Draize scores that correlated well with standard EpiOcular predicted Draize scores and historical in vivo Draize scores. HTS epidermal and ocular models such as these may thus help to lessen bottlenecks currently encountered in safety and efficacy testing of consumer products and topical therapeutics. This work was supported by NIEHS grant 5R44 ES010237-03.


10% fatty alcohol ethoxylate, 3-dimensional organotypic cultures, 96-well, Benzalkonium chloride, Centrimonium chloride, Cetyl pyridinium, Cocarride DEA, Cosmetics screen, Cytokine treatment, Disodium cocoamphodipropionate, Draize ocular, Draize scores, Drug permeation study, ET-50, Efficacy testing, EpiDerm, EpiDerm HST model, EpiOcular, EpiOcular HST, Epithelium, Glycerol, HTS, High content cell , High-throughput, High-throughput screening (HTS), Human epidermal, In vitro irritation/toxicity screen, Interlot culture, Intraplate culture, MTT, MTT assay, MTT tissue viability assay, Molecular biology assays, Normal human epidermal keratinocytes, OCL-200, Ocular epithelium, Preclinical safety, Robotic compatible, Safety and efficacy screen, Safety evaluation, Safety screening, Skin, Sodium cocoamphodipropionate, Sodium dodecyl sulfate, Sodium trideceth sulfate, Stearalkonium chloride, Surfactant blend, Topical therapeutics, Triton X-100

Materials Tested

Banzalkonium chloride, Sodium Chloride, Sodium Dodecyl sulfate, Triton X-100

Request a copy of this paper, click here.