ヒト神経細胞のIn vitro 3次元移植システムの開発と動的力学刺激付加による影響評価

抄録

In this study, by constructing a 3D transplantation system of human neurons in vitro, we aim to reproduce the process of neural fusion and find the possibility of application to intractable diseases of the nervous system such as Parkinson's disease and ALS with the aid of regenerative transplantation. In addition, we investigate how giving the vibration, which is one of the dynamic mechanical stimuli, affects this system. The system is constructed by gel-embedded culture of neuronal cells. Two neuronal cells colonies were sown with distance of 1400 to 1700 μm, and three-dimensional tissue formation between the two neuronal cells colonies was observed. A separately developed small piezo vibration stage was used to give vibration stimulation. The vibration conditions are with amplitude of 60 μm, frequency of 10 Hz, and of 8 hours vibration- 4 hours static repeated for 168 hours. Two patterns were conducted, one in which vibration is given only in the horizontal direction, and the other in which vibration is given every hour in the horizontal and vertical directions. As a result, using the "Neurolucida360", which is neuronal cells three-dimensional structural analysis software, it is confirmed that neuronal cells grow three-dimensionally, bind and fusion between recipient cells and donor cells. It is also confirmed that extension and sprout of neurites as well as fusion between neurites was promoted by giving the vibration stimulation.