摘要
A protocol for the isolation, purification and culture of motor neurons from newborn rat spinal cord was described and the effect of glial cell line-derived neurotrophic factor （GDNF） on the growth of neurite of motor neurons was investigated in vitro. Spinal motor neurons （SMNs） were dissociated from ventral spinal cord of postnatal day 1 rats. The culture system for SMNs was established by density gradient centrifugation, differential adhesion, and use of serum-free defined media and addition of exogenous GDNF. After 72-h culture, the cells displayed the characteristic morphology of motor neurons, exhibited extensive neuritic processes and were positive for choline acetyl- transferase （CHAT） expression. The neurite length of SMNs in GDNF groups was significantly longer than that in control group （P〈0.05）. This protocol can be adapted for various postnatal motor neurons studies.
A protocol for the isolation, purification and culture of motor neurons from newborn rat spinal cord was described and the effect of glial cell line-derived neurotrophic factor （GDNF） on the growth of neurite of motor neurons was investigated in vitro. Spinal motor neurons （SMNs） were dissociated from ventral spinal cord of postnatal day 1 rats. The culture system for SMNs was established by density gradient centrifugation, differential adhesion, and use of serum-free defined media and addition of exogenous GDNF. After 72-h culture, the cells displayed the characteristic morphology of motor neurons, exhibited extensive neuritic processes and were positive for choline acetyl- transferase （CHAT） expression. The neurite length of SMNs in GDNF groups was significantly longer than that in control group （P〈0.05）. This protocol can be adapted for various postnatal motor neurons studies.
基金
supported by a grant from National Research Program of Basic Medicine of China(No.2003CB515304)
