Fig. 3

Stable maintenance and superiority of NEL-MG on adult microglial genes despite the long-term passage culture. When NEL reached stratification, cells in one 25 T Flask were divided into two 25 T flasks, and then cultured for 21 days (d) for flow cytometry. A The ratio of CD11b⁺ microglia among NEL was approximately 50%, showing a stable maintenance ratio, despite the long-term passage culture (n = 3, mean ± SD) in flow cytometry. B NEL-MG were isolated based on the NEL passage culture (P0, P1, P2, P3, P4, P7, P8, and P10) and alterations in the adult microglial genes were examined compared to neonatal microglial cells. The experiment was performed three times independently (n = 3 per group) and the data represent mean ± standard error of the mean (SEM). A post hoc test was conducted using Dunnett’s multiple comparison test. *p < 0.05, ***p < 0.0001, ****p < 0.00001 compared to the 21-d group. Gray zone represents a value = 1 (neonatal microglia). C IBA-1, CX3CR1, and TMEM119 IRs and phagocytic function were not different in NEL-MG isolated from NEL at P0 (21d), P6 (90 d), and P10 (180 d). Scale bar = 50 μm