Table 3.
Effects of multiple neurotrophic factors on spinal motor neurons in short-term culture
| Survival (%) | |
|---|---|
| Two factors | |
| BDNF + GDNF | 46 ± 2 |
| BDNF + HGF | 48 ± 1 |
| FGF + BMP-7 | 38 ± 1 |
| BMP-4 + insulin | 45 ± 3 |
| Three factors | |
| BDNF + CNTF + HGF | 63 ± 2 |
| BDNF + FGF + GDNF | 66 ± 2 |
| FGF + BMP-7 + insulin | 65 ± 2 |
| Four factors | |
| FGF + HGF + BMP-7 + insulin | 75 ± 1 |
| CNTF + FGF + NT-3 + insulin | 72 ± 1 |
| BDNF + CNTF + GDNF + insulin | 75 ± 1 |
| Five factors | |
| BDNF + CNTF + GDNF + FGF + insulin | 80 ± 1 |
| GDNF + FGF + HGF + BMP-7 + insulin | 81 ± 1 |
| Six factors | |
| BDNF + CNTF + GDNF + FGF + BMP-7 + insulin | 89 ± 2 |
| BDNF + CNTF + GDNF + FGF + HGF + insulin | 84 ± 1 |
| Seven factors | |
| BDNF + CNTF + GDNF + FGF + HGF + BMP-7 + insulin | 87 ± 2 |
| Eight factors | |
| BDNF + CNTF + GDNF + FGF + HGF + BMP-7 + NT-3 + insulin | 91 ± 2 |
Approximately 3000 purified E15 spinal motor neurons cells were plated in triplicate in merosin-coated 96-well Falcon plates in 100 μl of L-15 medium without insulin, containing a plateau concentration in various combinations of the appropriate factor. After 3 d of culture, the percentage of cells surviving in each well was determined, using the MTT assay (see Materials and Methods; means ± SEM,n = 3, of a representative experiment is shown; all experiments were repeated at least three times).