大鼠胚胎脊髓神经细胞的原代培养及其神经营养素受体表达的初步研究

目的：建立哺乳动物胚胎脊髓神经细胞体外培养的细胞模型，提高培养用脊髓神经细胞的产率以及活性，使实验的细胞模型有良好的一致性和可比性。方法：18d的大鼠胚胎的脊髓经取材、分离、消化后作原代细胞培养，从细胞形态学及细胞计数角度观察细胞生长与分化过程。应用免疫细胞化学技术，观察了神经营养素受体TrKA、TrKB、TrKC在脊髓神经细胞的表达。结果：大鼠胚胎的脊髓神经细胞在体外条件下，可良好存活并有正常的表型分化，TrKA、TrKB、TrKC在细胞膜上呈阳性反应。其细胞数量(集落数)与形态以及存活周期可满足体外实验的基本要求。结论：这种分离培养方法简便、成功率高、稳定性强、细胞产量高且质量好，有助于离体研究的开展。

Initial Research of Primary Culture Embryonic Rat Spinal Cord Neurons and Its Expressions of Neurotrophic Receptors

Objective: To establish cytological model of spinal cord neurons in mammalian animals in vitro,and to increase the cell yield and cell viability of cultured rat's spinal cord to get a consistent and comparable laboratore cell model. Methods: The primary culture of spinal cord neurons carried out with SD embryonic rats aged 18 days, after being drew material,separated, and digested. The cells culture was observed with the methods of cytomorphology and cell counting. Immuno cell chemical technique was used to observe the expression of neurotrophic receptor, TrkA,TrKB and TrKC, on the spinal cord neuron. Results: The SD rat embryo spinal cord neuron could be cultured well in vitro, and had a normal differentiated appearence. TrKA, TrKB, and TrKC had their positive reactions on cellular membrane. The amount of cells (clones),morphology and survival cycle can meet the requirements of laboratory in vitro. Conclusion: The method reported is simple and convenient with high rate of success, and is steadiness with good output and better quality.