The Center for Alternatives to Animal Testing is an academic center affiliated with the Division of Toxicological Sciences in the Department of Environmental Health Sciences of the Johns Hopkins University Bloomberg School of Public Health.

Research Grants 2008-2009

3-D sertoli cell/genocyte co-culture system: in vitro model for developmental testicular toxicity

Xiaozhong Yu, MD, PhD
University of Washington

The development of in vitro models that monitor and which can identify altered testis development would provide important alternatives to in vivo testing. Such methods would allow for the assessment of reproductive and developmental effects induced by environmental agents and thus lead to significant refinement and reduction of in vivo animal use. Gonocytes exist in the neonatal testis and represent a transient population of male germ-line stem cells. It has been shown that stem cell self-renewal and progeny production is probably controlled by the neighboring differentiated cells and extracellular matrix (ECM) in vivo known as niches. Recently, we developed an in vitro three dimensional gonocyte/Sertoli cells co-culture (GSC) model with ECM, demonstrated this culture system creates an in vivo-like niche and supports germ-line stem cell functioning within a 3-D environment, permits the formation of a testicular-like multilayered architectural bio-structure that mimic in vivo characteristics of seminiferous tubules. We feel strongly that this novel in vitro GSC system will provide investigators with a simple, efficient, and highly reproducible alternative for the assessment of reproductive toxicity and the screening of testicular developmental toxicants. Thus, the purpose of this study is to further refine and characterize our in vitro 3-D GSC culture as a simple, efficient tool for screening testicular developmental toxicant by using a range of known in vivo male reproductive and developmental toxicants. This will be a systems based analysis using genomics and GO-quant technologies. We will also apply this 3-D GSC system to develop appropriate cell-based assays and biomarkers of susceptibility for testicular toxicity through the use of real-time nanotechnology biosensor assay methods. The establishment of a simple, efficient in vitro 3-D GSC culture model has the potential to greatly reduce the number of animals used through its ability to increase and refine the type of information obtained.