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本文研究了中脑多巴胺神经元在体外长期培养过程中的形态发育。动物选用胚胎14天SD大鼠,取中脑腹侧部制成细胞密度为2×10~5/ml的悬液,接种于24孔培养板中,培养1天至6周,以酪胺酸羟化酶免疫组化方法、单纯荧光组化方法和外源性儿茶酚胺荧光组化方法,观察中脑多巴胺神经元的体外长期培养发育状况。培养3天后多巴胺神经元已有单极或双极的突起发出,培养1周突起进一步伸长,并在沿途开始发出分支,有些末端见有生长锥。除散在分布的多巴胶神经元外,还能见到多巴胺神经元细胞群。培养2至3周,突起的分支增多,并明显出现念珠状膨大。在培养4-6周的标本中,仍可见到部分发育成熟的多巴胺神经元。多巴胺神经元在体外培养中的形态可分为二类,即梭形细胞和多极细胞。棱形细胞一般有两个突起,分别从胞体两极发出;多极细胞胞体呈圆形、多角形或三角形,可发出3-6个突起。实验结果表明多巴胺神经元的体外发育过程与在体相似。
This article studies the morphological development of midbrain dopamine neurons in long-term culture in vitro. Animals were selected embryos 14 days SD rats, ventral midbrain cells were made into a cell density of 2 × 10 ~ 5 / ml suspension, seeded in 24-well culture plates, cultured for 1 day to 6 weeks, with tyrosine Hydroxylase immunohistochemical method, simple fluorescence method and exogenous catecholamines fluorescence imaging method to observe the midbrain dopamine neurons in vitro long-term culture development. After 3 days of culture, dopaminergic neurons have emerged as unipolar or bipolar protrusions, which are further elongated after one week of culture and begin to branch off along the path and have growth cones at some ends. In addition to scattered dopa neurons, dopaminergic neuronal cell populations can be seen. After 2 to 3 weeks of culture, the number of raised branches increased and rosary enlargement clearly appeared. In 4-6 weeks of culture specimens, partially mature dopamine neurons can still be seen. The morphology of dopamine neurons cultured in vitro can be divided into two categories, that is, spindle cells and multipolar cells. Prismatic cells generally have two protrusions, respectively, from the bipolar body cell issue; polypoid cell body was round, polygonal or triangular, can issue 3-6 protrusions. The experimental results show that dopamine neurons develop in vitro similar to in vivo.