[Organotypic cerebral slice culture and the course of neuronal development in it].
OBJECTIVE
To establish a method of organotypic cerebral culture. So as to pave the way for building some neurodegenerative disease models.


METHODS
Organotypic cerebral cultures were prepared from prefrontal brain of neonatal SD rats. After culturing 7 to 14 days, 3 weeks, 4 weeks and 8 weeks, respectively, cerebral slices were fixed, dehydrated and sectioned in cryostat. The sections proceeded with Nissl staining and neurofilament high molecular weight (NFH) immunohistochemical staining. The difference was observed between controls and cultured slices using normal rats as controls.


RESULTS
Nissl staining showed that pyramidal neurons in cultured slices were increased in volume and lightened in staining. The delaminating construction was clear from 1 to 4 weeks after culturing. In cultured slices, immunohistochemical staining showed that NFH positive pyramidal cells appeared on layer V on the tenth day and on both layers V and III after culturing 12 days. In the control group, NFH positive pyramidal cells appeared on layer V in 5-day-old rats, and appeared on both layers V and III in over 3-week-old rats. In cultured cerebral slices, the number of pyramidal neurons on layer V in M1 area was invariable from 12 days to 2 months.


CONCLUSIONS
Orgaotypic cerebral culture can be used to study postnatal development for neocortex and build some in vitro models for neurodegenerative diseases.
Zheng
Mei
M
Department of Neurology, Peking University Third Hospital, Beijing 100083, China.
Fan
Dong-sheng
DS
Zhang
Jun
J
Song
De-mao
DM
Fan
Shao-guang
SG
D000818:Animals; D000831:Animals, Newborn; D004195:Disease Models, Animal; D005260:Female; D005625:Frontal Lobe / Q000166:cytology; D008297:Male; D009410:Nerve Degeneration; D009474:Neurons / Q000166:cytology*; D009924:Organ Culture Techniques / Q000379:methods*; D017966:Pyramidal Cells / Q000166:cytology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013687:Telencephalon / Q000166:cytology*