[Experimental study of primary culture methods for spiral ganglion cells of newborn rats].
OBJECTIVE
To optimize culture condition of spiral ganglion cells (SGCs) in vitro and to obtain highly purified SGCs.


METHODS
The spiral ganglions from newborn rats were digested in 0.25% trypsin and 0.001% DNase. The SGCs suspension was plated at a density of 10(9) cells/L. The cells were kept in serum free medium (DMEM/F12+B27 supplement). And 5 micromol/L cytosine arabinoside (Ara-C) was added to the medium on the second day and maintained for 48 hours. Serum medium with or without Ara-C was set as a control. The morphology and the purity of SGCs were observed under microscope.


RESULTS
Highly purified SGCs can be harvested in serum free medium (DMEM/F12+B27 supplement) with Ara-C. The SGCs were identified with mouse anti neurofilament protein antibody by immunohistochemistry methods.


CONCLUSIONS
The method used in this study is an optimal means to culture and purify SGCs that can meet the needs of further study.
Luo
Linghui
L
Department of Otolaryngology, Union Hospital, Huazhong University of Science and Technology, Wuhan, 430022, China.
Gong
Shusheng
S
Song
Peng
P
Yan
Kaisheng
K
D000818:Animals; D000831:Animals, Newborn; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D051381:Rats; D017207:Rats, Sprague-Dawley; D013136:Spiral Ganglion / Q000166:cytology*
D016895:Culture Media, Serum-Free