In Vitro & Alternative Toxicology

Non-Animal, In Vitro Testing Solutions.

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MB conducts Good Laboratory Practice (GLP) compliant toxicology assays as well as low-cost screening studies for clients in the cosmetic, consumer product, chemical, biotech and pharmaceutical industries.

In Vitro Sensitization Screening Assay

  • Using the 3D Reconstructed Human Epidermis protocol currently under review by ECVAM
  • Measures IL18 production in epidermal keratinocytes
  • For more information, see IVSA.

Ocular Irritation Assays

Bovine Corneal Opacity and Permeability Assay (BCOP)

  • Uses excised bovine corneas normally discarded
  • Measures two endpoints; 1) changes in opacity, reflecting protein denaturation and corneal injury, and 2) fluorescein permeability reflecting damage to corneal epithelium
  • Optional histological endpoint is available (H & E staining)
  • Measurement of pro-inflammatory mediators in liberated corneal cells (by flow cytometry or ELISA)

HET-CAM - Hen's Egg Test - Chorioallantoic Membrane (HET-CAM)

  • Uses fertilized chicken eggs
  • INVITTOX PROTOCOL Number 47

Chorioallantoic Membrane Vascular Assay (CAMVA)

  • Uses fertilized chicken eggs on embryonic day 10 or 14 of development
  • Measures hemodynamic effects, injury, and anti-angiogenic effects to the membrane microvasculature
  • Especially useful for alcohol-containing formulations

Porcine Cornea Reversibility Assay (PorCORA)

  • Uses excised porcine cornea normally discarded
  • Corneas maintained in culture for up to 21 Days
  • Measures "Days to Recover" after exposure to possible Irritants
  • Allows discrimination between Severe Irritant and Ocular Corrosive

Replacement Ocular Battery (ROBatt)

  • Uses tiered testing strategy of alternative ocular irritation assays (CAMVA, BCOP, PorCORA, PorFocal) to determine regulatory classification of a test material without the use of live animals.
  • Currently Under Development

In Vitro Toxicology Testing Assays

Three Dimensional Human Ocular Tissue Equivalent Systems

  • These models mimic characteristics of the epithelium of the eye
  • Unique properties of 3D models allow test articles to be applied topically, allowing testing of solids, organics, and insoluble materials

Keratinocyte/Fibroblast Viability Assay (Neutral Red or MTT)


MB Research uses normal human primary epidermal keratinocytes or fibroblasts (or mouse keratinocytes and fibroblasts)
  • Cells are representative of the eye and skin epithelium
  • Human cells are derived from neonatal, adolescent, or aged donors
  • Cytokine expression or release (LDH, PGE-2, interleukins) can be measured
  • Low serum or serum free protocol options are available

Keratinocyte Proliferation (turnover) Assay

  • Using primary human cell or mouse cell lines in serum-free or low serum media
  • Rate of proliferation or apoptosis is measured by vital dye uptake (MTT or neutral red) or by BrdU incorporation and flow cytometry

Dermal Irritation Assays

Three Dimensional Human Skin Equivalent Systems

The 3D models available at MB Research (MatTek EpiDerm™, SkinEthic systems and CellSystems EpiCS®) allow the application of the test material directly to the "skin" surface and permit the assessment of dermal toxicity via a wide variety of parameters.

  • Cytotoxicity via MTT, neutral red viability assay, or flow cytometry
  • Histological examination (H & E) of the epidermal layers, including the stratum corneum
  • Measurement of inflammatory mediator and lytokine release (PGE-2, nitric oxide, TNFa, IL1a, and other interleukins)
  • Dissolution of the tissue equivalent into a single cell suspension, with subsequent flow cytometric analysis allows evaluation of gene expression and peroxide production (antioxidant status of cells)

Dermal Corrosivity Assays


Corrositex™

  • An accepted alternative to the Draize rabbit corrosivity test that recently has been validated by ICCVAM for use for DOT, FHSA, CPSC, EPA and FDA testing
  • Allows Packing Group assignment of test materials
  • Can be used to compare the relative corrosivity of materials within Packing Groups for product development purposes. See www.corrositex.com for more information.

MatTek EpiDerm™ Corrosivity Test

  • Validated by ECVAM and ICCVAM as an in vitro alternative corrosivity test based on human skin cells
  • Allows testing of material not compatible with Corrositex™

Phototoxicity Assays

  • With 3T3 mouse fibroblast cells via the EC/COLIPA validated protocol. See www.3t3nru.com for more information.
  • Using 3D Tissue Construct protocols.
  • Human keratinocytes or fibroblasts

MB Research also performs other in vitro and alternative assays. Please refer to our Capabilities List or feel free to contact a Client Services Representative for assistance.