Abstract
Human iPS cells were established in 2007, and research on regenerative medicine has attracted worldwide attention since then. In 2017, it is scheduled to start clinical operation for the Parkinson's disease using neuronal progenitor cell differentiated from iPS cells. But, there are still some unsolved issues. First, there is not established a method to construct 3-D structures. Next, it takes very long time to induce differentiation of human iPS cells into neurons. Therefore, in this study, we constructed human neurons differentiated from human iPS cells in 3-D by using collagen gel. And we investigated the effect of continuous and interval dynamic stimulations in 3-D neuronal networking of human neurons. Dynamic stimulation（frequency: 15Hz, time: 24 hours, 48hours, 72hours, amplitude: 60μm）was applied using vibration stage to evaluate cell differentiation and proliferation efficiency. As a result, we succeeded in 3-D construction of human neuron networking, and establishing a method of gel-embedding culture. Furthermore, the differentiation growth rate was higher than the static culture in both continuous and interval culture. It was suggested that continuous stimulation is effective for 48 hours culture and 8:4 interval stimulation is effective for culture for 48 hours or more.
