Objective: To investigate the role of cytoskeleton in several important dynamic events during in vitro maturation of human oocytes. Study Design: Human germinal vesicle stage oocytes were divided randomly into control and study groups. After cultured for 24 hours, chromatin state and position, spindle formation and migration, cortical granules, and mitochondria distribution were evaluated. Results: In colchicine group, spindles did not form. Cortical granules migrated to the cortex but mitochondria maintained the peripheral distribution pattern in most of the oocytes. In cytochalasin B group, the migration of spindle and chromosomes to the cortex was prohibited. Microfilaments disruption influenced cortical granules migration but not redistribution of mitochondria. Conclusion: Meiosis progression could not go beyond metaphase I stage when microtubule or microfilament polymerization was prohibited in human oocytes. The migration of cortical granules to the cortex and redistribution of mitochondria to the inner cytoplasm were mediated by microfilaments and microtubules, respectively. © 2010 Mosby, Inc.