Laminin促进胚胎中脑多巴胺神经元生长发育的体外培养研究

在体外培养中研究了Laminin(LN)对大鼠胚胎中脑多巴胺神经元生长发育的影响。将胚胎(E15天)腹侧中脑细胞悬液接种于底部涂有LN的24孔培养板中,培养2—6天后取出。用酪氨酸羟化酶免疫组化ABC法观察多巴胺神经元的生长情况。培养48小时后见酪氨酸羟化酶免疫阳性细胞均有突起伸出,以双极细胞为主;有些突起分枝末端有活跃的生长锥,后者又司发出多根丝状伪足。在培养3到6天的标本中除见有单极和双极酪氨酸羟化酶免疫阳性细胞外,每孔均见有多极阳性细胞。这些阳性细胞的突起具有较丰富的分枝,突起长度多数长于胞体直径的6倍,最长的可达400μm以上。每孔酪氨酸羟化酶免疫阳性细胞数可达20到50个,并能见到群聚存在,与不含LN的对照组相比,在酪氨酸羟化酶免疫阳性突起的数量、长度及分枝等方面均有显著差异。实验表明LN可促进体外培养胚胎中脑神经元的生长发育及轴突延伸。

LAMININ PROMOTES GROWTH AND DEVELOPMENT OF EMBRYONIC MESENCEPHALIC DOPAMINERGIC NEURONS IN VITRO

The effects of laminin on growth and development of embryonic mesencephalic dopaminergic neurons were studied in vitro. Mesencephalic cells obtained from E15 rat were planted on 24 well culture plates previously coated with rat tail collagen plus laminin. Two to six days later, coverglass were examined with TH immunocytochemistry ABC method. After cultured for 48 hours, most of the TH positive cells had their processes extended. Some processes gave rise to terminal growth cones which ramified into several filopodia. By 3 to 6 days in culture, besides a few monopolar and some bipolar positive cells, multipolar positive cells were found in every well. The processes of these cells gave more branches. Most processes were even longer than six time of the diameter of their soma. The longest process was upto more than. 400μm in length. 20-50 positive cells and some small aggregates of DA neuron were found in each well. There was significant difference between the experimental group and control group in number, length and branches of their processes. Our results suggested that laminin can promote growth and development of mesencephalic neurons in vitro.