低密度体外培养大鼠海马神经元的形态学观察

Morphological observation of rat hippocampal neuron in low density culture in vitro

目的 :建立大鼠海马神经元低密度体外培养方法 ,并观察其发育分化过程中的形态学指征变化情况 .方法 :采用神经元与星形胶质细胞立体共培养方法 .首先以新生大鼠为实验材料培养纯化单层星形胶质细胞 ,随后以孕 18d胎鼠为实验材料 ,分离海马皮层 ,经胰酶消化制备单细胞悬液 .以(2～ 5 )× 10 3·cm-2 密度接种于包被有多聚赖氨酸的盖玻片上 ,而后采用无血清培养液与星形胶质细胞立体共培养 ,并对不同培养时间的神经元进行形态学观察 .结果 :利用低密度体外培养方法成功培养出海马神经元 ,其在不同发育分化阶段具有不同的形态学特征 .结论 :低密度培养的海马神经元不同发育分化阶段具有不同的形态学特征 。

AIM: To observe rat hippocampal neuron morphological characteristics at different developmental and differential stages in low density culture. METHODS: Neuron and astrocyte three dimensional co culture method was used. Acquire newborn rat astroglial monolayer and dissect embryonic 18 d rat hippocampus and obtain single cell suspension through trypsin digestion. Plate the suspension to poly L lysin coated coverglass in(2-5)×10 3 · cm -2 density. Transfer the coverglass to astrocyte culture section, co culture neurons with astrocytes in serum free medium and observe neuronal morphological characteristics at different stages. RESULTS: Rat hippocampal neuron in low density culture model was successfully acquired, which had distinct morphological characteristics at different stages. CONCLUSION: Hippocampal neuron in vitro has distinct morphological characteristics at different developmental and differential stages.