Objective: Selecting the most competent sperm is crucial for improving the success of intracytoplasmic sperm injection(ICSI). Chemotaxis-based sperm selection may facilitate the selection of spermatozoa with superior motility andfertilization capability. This study aimed to evaluate sperm selection based on chemotaxis induced by microvesicles(MVs) derived from follicular fluid (FF) and cumulus cells (CCs) using a simple microfluidic device.Materials and Methods: In this experimental study, FF and CCs were collected during ovum pick-up from 10 womenundergoing ICSI. MVs from both FF and CCs were isolated using the Exocib kit and characterized for particle size,polydispersity index, zeta potential, morphology, and protein content. To establish a chemoattractant gradient, MVs fromFF and CCs, as well as progesterone, were incorporated into agar plates. Nine experimental groups were studied: FF–derived MVs (FF-MV, 5, 25, and 50 μg/ml), CC–derived MV (CC-MV, 5, 25, and 50 μg/ml), FF (50 μg/ml), progesterone(0.03 μM), and a control (agar without chemoattractant). Sperm selection was performed in a microfluidic system usingsemen samples from 20 normozoospermic men. Sperm concentration, motility, progressive motility, viability, and DNAintegrity were analyzed following chemotaxis-based selection.Results: The isolated FF-and CC-derived MVs exhibited spherical morphology, a negative zeta potential, and werepolydispersed. The progesterone-containing plates showed sustained release. No significant differences were observedbetween the CC-MV and control groups in sperm parameters (P=0.630). Plates supplemented with 50 μg/mL of FF-MVmarkedly improved sperm concentration (P=0.002), progressive motility (P=0.010), and DNA integrity (P=0.001) comparedto the control group.Conclusion: FF-MVs, particularly at 50 μg/ml, possess chemoattractant properties and could selectively enrich spermpopulations with improved functional parameters. These findings highlight the potential of FF-MVs as a