High quality canine and rat Sertoli Cells available for researcher

Sertoli cells play critical roles in regulating spermatogenesis and testis development by providing structural and nutritional support. This study aimed to develop a standard protocol for canine Sertoli cell isolation and culture; and characterize its biological features, functionality, and application of compound toxicity testing. Canine testicles were received from the neuter clinic, and three-step of enzymatic digestion was applied to isolate Sertoli cells. We characterized the growth and purity of Sertoli cells with the expression of SOX9, GATA4, and Clusterin. In addition, we selected cadmium as a model toxicant to evaluate the toxic responses in the newly established Sertoli cells using High-content Analysis (HCA). With our optimized protocol, the purity of isolated Sertoli cells was above 95%, as determined with Sertoli cell-specific protein markers of SOX9 and GATA4. More importantly, primary Sertoli cell populations could be expanded rapidly in vitro, passaged (up to seven), and cryopreserved. The HCA-based assay revealed that cadmium at 1 μM induced both disruptions of cytoskeletal and DNA damage responses. Furthermore, we established an HCA assay with the newly isolated and optimized culture of canine Sertoli cells to evaluate the epigenetic markers of histone modification. We found cadmium-induced differential changes in histone modifications H3Me3K9, H3Me3K36, H4Me3K20, and H4acK5. In summary, we have established the standardized protocol to produce canine Sertoli cells with Sertoli cell-specific phenotype. The isolation and expansion of large quantities of canine Sertoli cells will provide broad applications in studying male infertility, reproductive toxicology, testicular cancer, and cell therapy.

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Mar 2023

Dr. Lei Yin delivered an invited talk entitled "High-content Imaging Analysis for Testicular Toxicity Prediction Using an 87-Compound Library in a Three-Dimensional Testis Model, --Building Integrated in vitro Models for Predicting in vivo Testing" at 40th Mountain West SOT Meeting at Santa Fe, NM (2022).

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Aug 2022

High-content Image based Analysis of Endocrine Disruptors

We published a munuscript titled "High-content analysis of testicular toxicity of BPA and its selected analogs in mouse spermatogonial, Sertoli cells, and Leydig cells revealed BPAF induced unique multinucleation phenotype associated with the increased DNA synthesis". The purpose of this study is to examine the selective toxicity of BPA, BPS), BPAF, and TBBPA on these testicular cells, including Sertoli, Leydig and spermatogonial cells. We developed a high-content image-based single-cell analysis and measured a broad spectrum of adverse endpoints related to the development of reproductive toxicology, including cell number, nuclear morphology, DNA synthesis, cell cycle progression, early DNA damage response, cytoskeleton structure, DNA methylation status, and autophagy. We introduced an HCA index and spectrum to reveal multiple HCA parameters and observed distinct toxicity profiling of BPA and its analogs among three testicular types. The HCA spectrum shows the dynamic, chemical-specific, dose-dependent changes of each HCA parameter. Each chemical displayed a unique dose-dependent profile within each type of cell. All three types of cells showed the highest response to BPAF at 10 μM across all endpoints measured. BPAF targeted spermatogonial cell more significantly at 5 μM. BPS more likely targeted Sertoli cell and Leydig cell and less at spermatogonia cells. TBBPA targeted spermatogonia, Sertoli cells, and less at Leydig cells. BPA is mainly targeted at Sertoli cells, followed by Ledig cells, and less at spermatogonial cells. Most importantly, we observed that BPAF induced a dose-dependent increase in spermatogonia cells, not in Sertoli and Leydig cells. In summary, our current HCA assays revealed the cell-type-specific toxicities of BPA and its analogs in different testicular cells. Multinucleation induced by BPAF, along with increased DNA damage and synthesis at low doses, could possibly have a profound long-term effect on reproductive systems.

Reprotox attended the New Mexico Humane conference on Aug 11, 2022. Our company has been working on developing novel in vitro models that promise to improve animal welfare and reduce the number of animals needed in future research. We are looking forward to meeting with New Mexico animal welfare advocates and finding collaboration opportunities.

SOT Poster Presentations in San Diego, March 27-31, 2022. We had three presentations at the Annual SOT meeitng, including "Development of Canine Testicular Organoids for Assessing Male Reproductive Toxicity", "High Content Imaging Analysis for the Testicular Toxicity Prediction Using an 87-Compound Library in a Three-Dimensional Testicular Cell Co-culture Model" and "Establishment, Characterization, and Toxicological Application of Canine Primary Sertoli Cells".

Reprotox attended NMBio’s second annual Knowledge Fair & Expo in October 2021. We enjoyed meeting with scientists, academic leaders, tech transfer specialists as well as entrepreneurs, business executives, venture capitalists and investors, and economic development professionals. We are looking forward to building strong relationships with industry colleagues in New Mexico.

Reprotox has been successfully relocated to Albuquerque New Mexico from Athens, GA. We are excited about this new improved R&D Lab space within the UNM Science and Technology Park.

WC10 (the 10th Congress on Alternatives and Animal Use in the Life Sciences) took take place at the Washington State Convention Center in Seattle on August 20-24, 2017. Dr. Yu co-chaired the Session of Models of Developmental and Reproductive Biology and presented "An animal-free In Vitro three-dimensional testicular cells co-culture model for evaluating male reproductive toxicants".