新生大鼠下丘脑神经元原代培养及形态学  

Primary culture of hypothalamic neuron and its morphological characteristics in newborn rat

作　　者：燕关玲 张华婧[1] 孙增荣[1] 张钰[1] 高娜[1] 李文[1] 

机构地区：[1]天津医科大学公共卫生学院劳动卫生与环境卫生学系,天津300070

出　　处：《解剖学杂志》2014年第1期34-36,71,共4页Chinese Journal of Anatomy

基　　金：国家自然科学基金（81273027）

摘　　要：目的：优化下丘脑神经元体外培养方法，观察神经元形态学特点及生长规律。方法：取SD新生鼠，剥离下丘脑，机械吹打法制成单细胞悬液，接种，24h后换为Neurobasal培养基；神经元特异性烯醇化酶（NSE）免疫细胞化学法计数阳性细胞；H-E染色、Nissl染色。结果：原代培养神经元6～8h内开始贴壁，培养5～7d突起增粗增长，连接成网，以多极神经元为主，胞体呈三角形、梭形、椭圆形；经NSE鉴定为神经元并且纯度约90％；细胞核大而圆，H-E染色胞核深蓝色，胞质红色，胞体可见尼氏颗粒。结论：改良后培养下丘脑的方法具有稳定的可重复性，可应用于下丘脑神经元培养实验及建立神经毒性研究模型。Objective： To optimize the techniques of primary cultivation of neurons in hypothalamus, and to determine the growth and morphological characteristics of the neuron. Methods： The hypothalamus of new born SD rats was dissected and dispersed mechanically with a Pasteur pipette, and then was cultivated. Neurobasal medium was added after 24 hours. Immunocytochemical method was used to count the percentage of the positive cells. And the cells were stained with H-E and Nissl staining. Results： Primary cultivated neurons were adhered in 6-8 hours. Neurites, being elongated, thicked and connected were found in 5-7days. Most neurons were multipolar, and cell bodies were triangle, spindle and ellipse in shape： The cells were green in NES staining. The purity degree of neurons was about 90%. Nucleus was blue by H-E staining and Nissl bodies were in the cytoplasm. Conclusion： This improved protocol is stable and repeatable, and it can be applied to cultivate the hypothalamus neuron and create the neurotoxicity model.

关 键 词：下丘脑神经元 原代培养 尼氏染色 

分 类 号：R322.811[医药卫生—人体解剖和组织胚胎学]