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Thiazovivin is a selective, cell-permeable small molecule that inhibits ROCK activity at a concentration 2 µM. Thiazovivin improves stemness maintenance of embryo-derived stem-like cells under chemically defined culture conditions in cattle. Thiazovivin directly targets Rho-associated kinase (ROCK) and increases expression of pluripotency factors. The process using thiazovivin could be easier, faster and more cost effective than transgene integration into somatic cells.