Bio-AlteR® 3D Testicular Cell Culture
The solution for reliable, robust and relevant spermatogenesis investigation in vitro
Bio-AlteR® is a novel 3D cell culture of seminiferous tubules enabling spermatogenesis in vitro.
Spermatogenesis in vitro
Bio-AlteR® is an innovative seminiferous tubules 3D cell culture technology enabling the spermatogenesis in vitro. The system mimics physiological conditions. It allows the study of both spermatogenesis mitotic and meiotic phases, as well as the first steps of spermiogenesis. The technology preserves the original 3D structure of the seminiferous tubules, and the Sertoli cells remain polarized which is crucial for the differentiation of germ cells. The blood testis barrier is maintained, which allows the study of drug permeation across the barrier. Thus, this technology bridges the gap between in vitro to the in vivo techniques.
One well, two compartments
Cells are cultured in a bicameral chamber. This system allows the study of the impact of compounds (substances, drugs or molecules) added into the basal compartment on both the blood testis barrier integrity and on differentiating germ cells. The basal compartment mimics the blood compartment whereas the apical compartment reproduces the seminiferous tubules lumen environment. The cell culture media can be analyzed to identify product release and explore biomarkers produced as a result of toxicity.
Bio-AlteR® has proven its efficacy and reliability for the testing of many molecules such as endocrine disruptors, heavy metals, pesticides, pollutants, others… [Download the list of compounds tested by Bio-AlteR®]
Experimental protocol advantages
- Up to 4 weeks’ time duration of drugs/ chemicals testing
- Cells grow in a serum free medium
- Ability to test the substance on differentiating germ cells
- Drugs/Chemicals are added in the basal compartment to mimic the blood compartment
- Possibility of testing low concentrations of toxicants ("physiological")
- Capacity to evaluate the drugs/chemicals toxic effect reversibility
Our approach: Testicular "Physio-toxicology"
- Test the effects of a toxicant on differentiating germ cells
- Test concentrations of toxicants « close to » concentrations found in body fluids (including identified metabolites)
- Test the effects on relevant « end points »
- In every instance « think of physiology »
- Test the effects of a toxicant at different times of development, if necessary
- If using in vitro approaches, check how these models relate to the in vivo (physiological) situation
- In order to prevent "false negative" results experimental models must allow testing concentrations of toxicants close to those found in biological fluids (Geoffroy-Siraudin et al., 2010; 2012; Perrard et al., 2013).
- Also, in order to prevent "false positive" results, particular attention should be given to the concentrations of toxicants which are experimentally tested, and do not work with “excessive” doses.
Modification of cell number population
- Cell viability
- Percentage evaluation of six main cell population
Blood testis barrier integrity
- Trans-epithelial resistance (only in rats) [Download TEER measurement description]
- mRNA or protein expression of blood testis barrier components: Connexin 43, Claudin, N-Cadherin…
Determination of a meiotic index
Analysis of specific cell gene expression
- RT-qPCR six main cell populations and blood testis barrier components
Peptide profiling of the culture supernatant/cultured cells
Cross-species comparison studies are possible using Bio-AlteR® to better understand the mechanism(s) involved in a testicular toxic effect.