Figure 4.

Survival of CTB-immunopurified CSMNs in vitro. A, Percentage of purified P3 CSMNs surviving after 5 DIV in the various media listed. Neg, Basal serum-free medium (see Materials and Methods); CPT-cAMP, CPT-cAMP added to basal medium; all remaining single/multiple factors were added in the presence of CPT-cAMP; All, CXCL12 plus IGF2 plus PTN plus BDNF with or without IGF1 (no significant differences noted with/without IGF1 when the other four factors were present); ECM, fresh 10×ECM added at plating and day 3 feeding. All data presented were normalized to negative controls (Neg or CPT-cAMP) (±SEM). *p < 0.001, multiple pairwise ANOVA post hoc Student–Newman–Keuls test; **p < 0.0001, two-tailed Student's t test; all statistical differences were found with control CPT-cAMP, no significant differences were detected between the various factor/ECM conditions. Neg, n = 20; CPT-cAMP, n = 41; BDNF, n = 33; IGF1, n = 21; IGF2, n = 33; PTN, n = 26; CXCL12, n = 15; All, n = 19; ECM, n = 11; ECM+All, n = 13; for each “n” value, 50–300 cells per well were counted. B1, B2, Insulin+BDNF supports short-term but not long-term CSMN survival. Two independently performed assays of purified P3 CSMN survival at various DIV in media containing CPT-cAMP and insulin, with or without BDNF are shown. All data are ±SEM (n = 3). C, Survival of purified P7 CSMNs 1–3 DIV in media containing CPT-cAMP and insulin, with or without BDNF.