Ziętek Marta Marlena, Bihorac Ajna, Wenta-Muchalska Elżbieta, Duszewska Anna Maria, Olech Wanda, Sampino Silvestre, Bernat Agnieszka
Department of Experimental Embryology, Institute of Genetics and Animal Biotechnology Polish Academy of Sciences, 05-552 Jastrzębiec, Poland.
Department of Morphological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, 02-787 Warszawa, Poland.
Int J Mol Sci. 2025 May 2;26(9):4327. doi: 10.3390/ijms26094327.
The European wisent (), an iconic yet genetically vulnerable species, faces ongoing conservation challenges due to a restricted gene pool. Advances in induced pluripotent stem cell (iPSC) technology offer promising prospects for preserving and restoring genetic diversity in endangered species. In this study, we sought to reprogram wisent somatic cells into iPSCs using the PiggyBac transposon system, a non-viral method known for being successfully applied in bovine species. We applied a six-factor reprogramming cocktail (, , , , , ) alongside small-molecule enhancers to fibroblasts isolated from adult wisent tissue. While initial colony formation was observed, the reprogrammed cells exhibited limited proliferation and failed to maintain stable pluripotency, suggesting intrinsic barriers to complete reprogramming. Despite optimizing culture conditions, including hypoxia and extracellular matrix modifications, the reprogramming efficiency remained low. Our findings indicate that wisent somatic cells may require alternative reprogramming strategies, such as new-generation delivery systems and epigenetic modulators, to achieve stable iPSC lines. This study underscores the need for species-specific optimization of reprogramming protocols and highlights the potential of emerging cellular technologies for conservation efforts. Future research integrating advanced reprogramming tools may pave the way for genetic rescue strategies in wisent and other endangered species.
欧洲野牛（）是一种具有标志性但基因易受影响的物种，由于基因库有限，面临着持续的保护挑战。诱导多能干细胞（iPSC）技术的进步为保护和恢复濒危物种的遗传多样性提供了有希望的前景。在本研究中，我们试图使用PiggyBac转座子系统将欧洲野牛体细胞重编程为iPSC，这是一种已知已成功应用于牛物种的非病毒方法。我们将一种六因子重编程鸡尾酒（，，，，，）与小分子增强剂一起应用于从成年欧洲野牛组织中分离的成纤维细胞。虽然观察到了最初的集落形成，但重编程细胞的增殖有限，并且未能维持稳定的多能性，这表明完全重编程存在内在障碍。尽管优化了培养条件，包括缺氧和细胞外基质修饰，但重编程效率仍然很低。我们的研究结果表明，欧洲野牛体细胞可能需要替代的重编程策略，如新一代递送系统和表观遗传调节剂，以获得稳定的iPSC系。本研究强调了对重编程方案进行物种特异性优化的必要性，并突出了新兴细胞技术在保护工作中的潜力。整合先进重编程工具的未来研究可能为欧洲野牛和其他濒危物种的基因拯救策略铺平道路。
