Application of centromeric fluorescence in situ hybridization probes for embryological and cell studies—A practical approach to validate chromosome stability in Bovinae

BACKGROUND: Advances in reproductive and cellular biotechnology have significantly enhanced the ability to study, conserve, and manipulate species within the Bovinae family, including domestic cattle and the endangered European bison (wisent). Central to the success of such technologies, including in vitro fertilization, somatic cell nuclear transfer, and the culture of pluripotent stem cells (PSCs), is the chromosome stability of cultured cells and embryos. RESULTS: Here, we present the design and application of a specific centromere Cot-1 DNA probe for fluorescence in situ hybridization (FISH). We demonstrate its utility in assessing chromosomal stability of bovine and wisent somatic cells and in vitro-produced embryos. The probe shows high specificity for Bovinae cells and can be used on both metaphase plates and interphase nuclei, enabling rapid cytogenetic analysis even in slow-dividing or senescent cells. CONCLUSIONS: The newly generated Cot-1 DNA-based FISH probe provides a rapid and reliable method for species-specific cytogenetic monitoring in Bovinae. Furthermore, it allows accurate monitoring of chromosomal stability and quality control in PSC systems reliant on feeder layers and offers a streamlined approach for monitoring embryo chromosomal integrity in vitro. Broader application of this method may improve outcomes in reproductive technologies, conservation programs, and the generation of stable cell lines for biotechnological and agricultural use.