[Objectives] Animal cell culture technologies become more crucial in ecotoxicological researches. Due to some technical difficulties, studies on tissular cell culture of freshwater mussels are still limited. Although primary culture of gill cell in Anodonta woodiana was succeeded in previous study, breakthroughs have not yet been obtained for other tissues. Therefore, we intend to expand the primary culture of A. woodiana cells from four tissues (hemocyte, digestive gland, mantle, and foot). [Methods] The healthy 2-year-old “standardized” A. woodiana were used to obtain tissue cells. Different culture conditions (e.g., culture media, temperature) were designed to comparatively reveal survival rates and cell viabilities of the above four tissular cells so as to optimize the culture conditions for the primary culture. [Results] The results showed that the survival rates of the above four tissular cells were affected by different culture temperature, time and medium type. The survival rates were 94.67% ± 0.47% in culture medium 2 at 20 ℃ for 96 h for the hemocytes, 93.67% ± 1.70% in medium 3 at 20 ℃ for 48 h for the digestive gland cells, 93.67% ± 1.70% in medium 1 at 15 ℃ for 48 h for the mantle cells, and 94.33% ± 0.94% in medium 1 at 15 ℃ for 48 h for the foot cells (Fig. 2). Compared to 25 ℃, the survival rates of four tissular cells were higher at 20 ℃ and 15 ℃ for 96 h. [Conclusion] The study obtained the breakthrough on primary culture of hemocyte, digestive gland, mantle, and foot cells in A. woodiana. The study provides basic data for the establishment of cell lines of A. woodiana. It can also provide theoretical support for subsequent studies of toxicology at the cellular level of freshwater mussels in the future.