Fig. 1

Tri-culture media is capable of supporting neurons, astrocytes, and microglia. a Representative fluorescence images of the tri- and co-cultures at DIV 7 and 14. The cultures were immunostained for the three cell types of interest: neurons—anti-βIII-tubulin (red), astrocytes—anti-GFAP (green), microglia—anti-Iba1 (orange), general nuclear stain DAPI (blue). Microglia are present in the tri-culture at both time points but are absent in the co-culture. Scale bar = 100 μm. b Quantification of the number of each cell type and total number of cells per mm² for each condition shown in Fig. 1a (mean ± SD, n = 3). c Percent of total cell population represented by each cell type in co-culture vs. tri-culture conditions. d Quantification of the percent area coverage of the neurons and astrocytes in the co- and tri-cultures at DIV 7 and 14 (mean ± SD, n = 3). b, d The letters above the bars indicate statistically distinct groups (p < 0.05), while the points indicate the values of the technical replicates