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In recent years, significant improvements have been made in in vitro oocyte maturation methods in many mammalian species, including pigs. Despite using morphological selection as well as creating an environment that is very similar to in vivo conditions, the obtained in vitro matured oocytes still display a lower quality than oocytes obtained in vivo. Methods for in vitro maturation of pig oocytes have been significantly improved in recent years, resulting in effectiveness of about 50–90% of oocytes reaching the Met II stage (Nguyen et al., 2017; Chen et al., 2020). This is why attempts are being made to modify maturation conditions, with the goal of producing mature in vitro porcine oocytes similar to those obtained in vivo. Thymosin (TMS) is a peptide hormone first described following its discovery by Goldstein (Goldstein et al., 1966). Apart from its immunomodulatory functions, TMS promotes angiogenesis and prevents inflammation, apoptosis and tumorigenesis (Li et al., 2010; Kumar and Gupta, 2011). It demonstrates regenerative effects such as stimulation of hair regrowth, acceleration of collagen production, healing of wound burns and dermal damage and heart muscle regeneration (Tseng et al., 2002; Hinkel et al., 2015). To date, the regenerative qualities of TMS have been tested only on somatic cells and bovine oocytes (Skowronek et al., 2016; WierzchośHilczer, 2017). Taking into consideration TMS regenerative qualities, it was assumed that utilizing TMS for porcine oocyte maturation – especially oocytes with a lower morphological quality – would result in improved quality of the oocytes and developing embryos. So, the objective of this study was to determine the effect of TMS on the in vitro maturation of pig oocytes and on in vitro fertilization (IVF), which would be used to assess the quality of obtained embryos. ABSTRACT. Thymosin (TMS) is a peptide hormone that has regenerative qualities. To date, the regenerative qualities of thymosin have been tested only on somatic cells and bovine oocytes. The objective of this study was to determine the effect of thymosin on the in vitro maturation of pig oocytes and on in vitro fertilization (IVF), which would be used to assess the quality of obtained embryos. Pig oocytes (n = 463) were matured in vitro to the Met II stage in medium supplemented with TMS. After IVF the zygotes were cultured in vitro until they reached the blastocyst stage. The number of cells and the level of nuclear DNA fragmentation were evaluated. The addition of TMS to the maturation medium resulted in a significant increase in the percentage of competent oocytes that, after fertilization, developed to the blastocyst stage. Moreover, supplementing the maturation medium with TMS had a positive effect on the quality of obtained blastocysts by limiting the occurrence of apoptosis. Received: 5 May 2020 Revised: 5 October 2020 Accepted: 19 November 2020

Effect of thymosin β on maturation of pig oocytes and quality of <i>in vitro</i> produced embryos