| Summary: | The purpose of this study was to determine the impact of intraovarian injections of a reconstituted lyophilized growth-promoting factor extracted from horse blood platelets (L-GF<sup>equina</sup>) on the number of ovarian follicles, the recovery of cumulus–oocyte complexes (COCs), and embryo development to the blastocyst stage in Holstein cows. Thus, 12 Holstein cows were assigned to three protocols. According to the number of punctured follicles in protocol 1, ovum pick-up (OPU) was conducted on days 6 and 14 of the cycle (day 0 = estrus). In protocol 2, every large follicle (more than 7 mm) was removed, and 1 mL of L-GF<sup>equina</sup> was intraovarian injected (day 0). Two days later, equine chorionic gonadotropin (eCG) was administered, and OPU sessions were conducted on days 6, 10, and 14. The same ovarian stimulation procedure as that in protocol 2 was performed in protocol 3, except that equine L-GF<sup>equina</sup> was not supplied. OPU was carried out on days 6 and 10 of the cycle. The results indicate that the intraovarian injection of L-GF<sup>equina</sup> significantly (<i>p</i> < 0.05) increased the number of OPU sessions per cycle, the recovery of cumulus–oocyte complexes (COCs), and the production of blastocysts. In conclusion, an intraovarian injection of L-GF<sup>equina</sup> can improves OPU-IVEP results in Holstein cows.
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