Development of a In vitro 2-D transplantation system of human neurons and the effects of dynamic stimulation on the system

Abstract

In this study, we develop 2-D in vitro transplantation system for human neurons. Dynamic stimulation was added for the purpose of promoting the connection between neural networks and their effects were evaluated. First, differentiation of human iPS cells was induced into motor neural progenitor cells. After 35 days, the neural progenitor cells are adhered, and are differentiated into neurons colony. Then 24 hours after adhesion, new neural progenitor cells colony are seeded near by as transplanted donor cells. Thus, transplantation was simulated by shifting the timing to start the adhesion culture. After transplantation and confirmation of cells adhesion, dynamic stimulation was applied. In this study, we established a living fluorescent staining method using NeuroFluor NeuO. Using this method and conventional method, staining was performed on day 1, 3, 5 and 7 after transplantation, and evaluation was performed. As a result, it was confirmed that the application of the dynamic stimulation promoted the differentiation growth rate over entire period, and also the networking between the transplanted neuronal donor cells colonies and the recipient ones was promoted by the dynamic stimulation.