Hatanaka H, Nishio C, Kushima Y, Tsukui H
Division of Protein Biosynthesis, Institute for Protein Research, Osaka University, Japan.
Neurosci Res. 1990 Jun;8(2):69-82. doi: 10.1016/0168-0102(90)90060-r.
We have established a primary neuronal cell culture technique from the postnatal (P11 to P15) rat CNS to study the nerve growth factor (NGF) response to basal forebrain cholinergic neurons. The survival of septal cholinergic neurons in culture was monitored both by the determination of choline acetyltransferase activity and by counting acetylcholinesterase-positive cells. Cells obtained from postnatal septal regions were found to require a plentiful oxygen supply during the dissociation of the cells. NGF-mediated survival of the septal cholinergic neurons was similarly observed in the cultures by using different plating cell densities up to 12.5 X 10(5) cells/cm2. These results suggest that the promotion by NGF of cell survival in culture is independent of plating cell density.
我们建立了一种从出生后(P11至P15)大鼠中枢神经系统获取原代神经元细胞培养技术,以研究神经生长因子(NGF)对基底前脑胆碱能神经元的反应。通过测定胆碱乙酰转移酶活性和计数乙酰胆碱酯酶阳性细胞来监测培养中隔区胆碱能神经元的存活情况。发现从出生后隔区获得的细胞在解离过程中需要充足的氧气供应。通过使用高达12.5×10(5)个细胞/cm2的不同接种细胞密度,在培养物中同样观察到NGF介导的隔区胆碱能神经元存活。这些结果表明,NGF对培养中细胞存活的促进作用与接种细胞密度无关。