Establishment of a cartilage tissue regeneration technique is needed to treat bone diseases such as osteoarthritis. However, problems such as necrosis of cells due to high-density cell culture and shear stress by gravity have not yet been solved. Thus, we examined an RWV bioreactor that simulates a micro-gravity environment. Rabbit bone marrow cells were seeded in the RWV reactor. Large cell aggregate with a longer diameter of about 1.5 cm was formed after 4 weeks. Histochemical and biochemical analysis confirmed that large and homogenous three-dimensional cartilage tissues were successfully generated without necrosis by culture of bone marrow cells in an RWV bioreactor.