Keywords

vasculogenesis, developmental toxicology, in vitro, type I collagen, oligomer

Presentation Type

Event

Research Abstract

In vivo toxicology assays are expensive, low-throughput, and often not predictive of a human response. Three-dimensional in vitro human cell-based tissue systems incorporating cell-cell and cell-matrix interactions have promise to provide high-throughput, physiologically-relevant information on the mechanism of the toxin and a more accurate assessment of the toxicity of a chemical before progression to human trials. Quantification of the disruption of vasculogenesis, the de novo formation of blood vessels from endothelial progenitor cells, can serve as an appropriate indicator of developmental toxicity since vasculogenesis is critical to the early development of the circulatory system. The current routinely used in vitro angiogenesis assays are 2D and thus not physiologically-relevant and analyzed semi-quantitatively. Recently, a 3D in vitro model of vasculogenesis, involving a type I collagen oligomer matrix and endothelial colony forming cells, was described to be capable of producing and maintaining lumenized blood vessel networks up to 14 days. Here we utilized this model to develop a 3D vasculogenesis assay. We quantified the performance of this assay with a set of known angiogenesis modulators (imazamox, thalidomide, 2-methoxyestradiol, levamisole) through cytotoxicity testing and 3D image analysis. Preliminary data suggests a difference in the drug sensitivity and response when measured with the 3D vasculogenesis assay and compared to a well-established 2D angiogenesis assay. We expect our model to more closely mimic results from in vivo studies, but further validation is needed. This new 3D vasculogenesis model can provide a physiologically-relevant, high-throughput screening and mechanistic assay for applications in pharmaceutical development and developmental toxicology.

Session Track

Health

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Aug 7th, 12:00 AM

Tissue Engineering: Applications in Developmental Toxicology

In vivo toxicology assays are expensive, low-throughput, and often not predictive of a human response. Three-dimensional in vitro human cell-based tissue systems incorporating cell-cell and cell-matrix interactions have promise to provide high-throughput, physiologically-relevant information on the mechanism of the toxin and a more accurate assessment of the toxicity of a chemical before progression to human trials. Quantification of the disruption of vasculogenesis, the de novo formation of blood vessels from endothelial progenitor cells, can serve as an appropriate indicator of developmental toxicity since vasculogenesis is critical to the early development of the circulatory system. The current routinely used in vitro angiogenesis assays are 2D and thus not physiologically-relevant and analyzed semi-quantitatively. Recently, a 3D in vitro model of vasculogenesis, involving a type I collagen oligomer matrix and endothelial colony forming cells, was described to be capable of producing and maintaining lumenized blood vessel networks up to 14 days. Here we utilized this model to develop a 3D vasculogenesis assay. We quantified the performance of this assay with a set of known angiogenesis modulators (imazamox, thalidomide, 2-methoxyestradiol, levamisole) through cytotoxicity testing and 3D image analysis. Preliminary data suggests a difference in the drug sensitivity and response when measured with the 3D vasculogenesis assay and compared to a well-established 2D angiogenesis assay. We expect our model to more closely mimic results from in vivo studies, but further validation is needed. This new 3D vasculogenesis model can provide a physiologically-relevant, high-throughput screening and mechanistic assay for applications in pharmaceutical development and developmental toxicology.