INTRODUCTION: Early preimplantation embryos of mammals exhibit pronounced sensitivity to hyperosmotic conditions, which results in an embryonic developmental block. The reduction of medium osmolarity or the supplementation with organic osmolytes can eliminate this arrest. Therefore, cell volume homeostasis is essential for embryonic development METHODS: The IVM of porcine oocytes was divided into two stages, i.e. the first 22 h as the first stage, and the last 22 h as the second stage. In the presence of 1 mM glycine, we adjusted the osmolality of the culture medium from low to high (290 mOsM for the first 22 h and 320 mOsM thereafter) by adding raffinose, which cannot be used by animal cells, in a culture stage-dependent manner. RESULTS: Stage-dependent adjustment of simplified medium PZM-3 osmolarity (290 mOsM for the first 22 h and 320 mOsM thereafter) in the presence of 1 mM glycine significantly improved the quality of porcine oocyte maturation CONCLUSION: Our study demonstrates that optimizing cell volume regulation can further enhance the developmental potential of oocytes cultured