Figure 5.

ACM and MCM facilitate MN maintenance in matured MN cultures deprived of trophic support. A, MNs were cultured with 20 μg/ml ACM, MCM, or MEx for 2 d. After 2 DIV, some cultures were counted, whereas the others were rinsed and were replaced with fresh media containing no additional trophic support. The remaining cells were counted 3 d later at 5 DIV. Data are expressed as the percentage of cells present at 2 DIV that remain at 5 DIV (*p < 0.05, ***p < 0.001; ANOVA with Tukey–Kramer post hoc test; n = 8). B, MNs were cultured from plating with 20 μg/ml ACM. At 2 DIV, cultures were counted, rinsed thoroughly, and replaced with fresh media containing 20 μg/ml of either ACM or MCM. At 5 DIV, cells were counted to determine the number of MNs remaining. Quantification of cell counts described in A indicate that both ACM and MCM are able to maintain MNs to an equivalent degree and that the death-inducing factor (s) present in MCM is incapable of exerting its function under these culture conditions. C, Representative images of the conditions described in B depict calcein AM-labeled MNs 3 DIV after media were changed. Images were captured at 20×.