Culture of motor neurons from newborn rat spinal cord.
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 acetyltransferase (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.
Cheng
Shigang
S
Department of Urology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China. chenggang527@163.com
Shi
Ying
Y
Hai
Bo
B
Han
Xiaomin
X
Chen
Zhaohui
Z
Li
Bing
B
Xiao
Chuanguo
C
D000818:Animals; D000831:Animals, Newborn; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000494:pharmacology*; D009046:Motor Neurons / Q000166:cytology*; D016501:Neurites / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000166:cytology*
D016895:Culture Media, Serum-Free; D051100:Glial Cell Line-Derived Neurotrophic Factor