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Objectives: To observe the growth of Gardnerellavaginalis biofilm in vitro and the effects of Lactobacilli on the formation of Gardnerellavaginalis biofilm.Methods: We cultivated GardnerellavaginalisATCC14018 in vitro to form the biofilm and assessed the amount and the morphology of biofilm at 4h, 8h, 12h, 16h, 20h, 24h, 48h, 72h, 96h and 7th day respectively.Then, we added equal amount of Lactobacilli at 0h, 24h, 48h after the biofilm had formed and co-cultivated these bacteria for another 24h to observe the formation of Gardnerellavaginalis biofilm with LIVE/DEAD BacLight Bacterial Viability Kit.The five strains of Lactobacilli were isolated, purified and identified by the laboratory of gynecology in Peking University First Hospital from the vaginal secretes of health women.Result: The OD values were significantly different at various time points by micro-quantitative analysis.Scanning electron microscope images showed that the scattered single colony existed at 4~20h and began to aggregate at 24h.The images also showed that the colonies attached mutually were surrounded by extracellular polymeric substance at 48h, but there were no changes after 48h.By fluorescent staining, we observed that biofilm became thinned and dispersed at 0h, 24h after adding Lactobacilli.However, there were no differences in density of the colonies.The number of dead bacteria increased and Lactobacilli were the major live bacteria at 48h.Conclusion: Our results suggest that the process of biofilm formation is relatively slow.Gardnerellavaginalis begin to form biofilm at 24h after cultivation and the biofilm come to be mature after 48h approximately.Lactobacilli can also interfere the formation of biofilm, and different species of Lactobacilli have various effects on the biofilm when in different periods of formation.