The aim of this study was to evaluate the improvement of testicular sperm motility following different culture conditions such as human follicular fluid (hFF) and temperature. Testicular tissues obtained from azoospermia (n=21) were minced into small pieces by blade and recovered sperm suspension were cultured in Ham's F10 with or without 40% hFF at different temperatures (Group I: 37/with hFF, Group II: 32/withGroup III: 37/without, Group IV:32 /without The motility and viability of sperm were monitored during culture for 48 hours. Initial motility of testicular sperm was 10.91.9%. After 24 hours culture, sperm motility was 23.52.1% (Group I), 8.11.1% (Group II), 10.4 1.4% (Group III) and 4.00.8% (Group IV), respectively. After 48 hours, the motility had been changed as 322.3% (Group I), 14.31.7% (Group II), 5.3 1.4% (Group III) and 4.30.9% (Group IV). In hFF group (I and II), sperm motility of group I cultured at 37 was higher than those of group II at 32. But, sperm viability of group I cultured at 37 was lower than those of group II at 32 (54.44.1% vs. 59.43.7%) after cultured for 48 hours. We acquired the best motility of testicular sperm when performed in vitro culture for 48 hours in hFF supplemented medium at 37. Increase of sperm motility by in vitro culture could be useful tool fur human TESE-ICSI program.