ESTABLISHMENT OF PURE NEURONAL CULTURES FROM FETAL-RAT SPINAL-CORD AND PROLIFERATION OF THE NEURONAL PRECURSOR CELLS IN THE PRESENCE OF FIBROBLAST GROWTH-FACTOR
ESTABLISHMENT OF PURE NEURONAL CULTURES FROM FETAL-RAT SPINAL-CORD AND PROLIFERATION OF THE NEURONAL PRECURSOR CELLS IN THE PRESENCE OF FIBROBLAST GROWTH-FACTOR
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DOI：
10.1002/jnr.490290410
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发表时间：
1991-08-01
影响因子：
4.2
通讯作者：
SENSENBRENNER, M
中科院分区：
文献类型：
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作者：
DELOULME, JC;BAUDIER, J;SENSENBRENNER, M
一个几乎纯的神经元细胞的原代培养系统，从14日龄的胎鼠脊髓已开发相结合的preplating步骤，使用化学成分确定的无血清培养基，和硼酸化的聚赖氨酸包被的盘子，防止形成细胞聚集体。发现约98%的细胞被神经元特异性烯醇化酶抗体免疫染色，证实了它们的神经元性质。文化基本上是由一个人口的非运动神经元，并含有很少的运动神经元，其特点是他们的大尺寸和多极方面，乙酰胆碱酯酶（AChE）的存在下，和强烈的免疫反应生长相关蛋白GAP-43。神经元前体细胞也存在于这些培养物中，并在前3天增殖。牛脑碱性成纤维细胞生长因子（bFGF）的加入刺激它们的增殖超过2天的时间，通过测量[I-125]碘脱氧尿苷掺入和溴脱氧尿苷掺入细胞核后的免疫细胞化学反应测定。神经元特异性烯醇化酶免疫组化染色显示增殖细胞为神经元。重组人碱性成纤维细胞生长因子和牛脑酸性成纤维细胞生长因子（aFGF）发挥类似的作用。其他生长因子，包括表皮生长因子（EGF），转化生长因子β-1（TFG-beta-1）和凝血酶，对这些神经元细胞的增殖活性没有影响。bFGF对运动神经元的存活和整个神经元群体的纤维生长没有影响。然而，碱性成纤维细胞生长因子影响双极乙酰胆碱酯酶阳性神经元的发展，可能属于非运动神经元人口。这些数据表明，碱性成纤维细胞生长因子和碱性成纤维细胞生长因子是有丝分裂原从大鼠脊髓中的文化和碱性成纤维细胞生长因子的影响，是乙酰胆碱酯酶阳性的脊髓神经元的亚群的发展。
A primary culture system of nearly pure neuronal cells from 14-day-old fetal rat spinal cord has been developed by combining a preplating step, the use of a chemically defined serum-free medium, and borated polylysine-coated dishes that prevented the formation of cell aggregates. About 98% of the cells were found to be immunostained with neuron-specific enolase antibodies, confirming their neuronal nature. The cultures are composed essentially of a population of non-motoneurons and contain few motoneurons, characterized by their large size and multipolar aspect, the presence of acetylcholinesterase (AChE), and the intense immunoreaction for growth-associated protein GAP-43. Neuronal precursor cells are also present in these cultures and proliferate during the first 3 days. The addition of bovine brain basic fibroblast growth factor (bFGF) stimulates their proliferation over a period of 2 days, as determined by measurement of [I-125]iododeoxyuridine incorporation and by immunocytochemical reaction after bromodeoxyuridine incorporation into nuclei. The proliferating cells were characterized as neurons by immunostaining against neuron-specific enolase. Recombinant human bFGF and bovine brain acidic FGF (aFGF) exerted similar effects. Other growth factors, including epidermal growth factor (EGF), transforming growth factor beta-1 (TFG-beta-1), and thrombin, were without effect on the proliferative activity of these neuronal cells. bFGF has no effect on the survival of motoneurons and on the fiber outgrowth of the whole neuronal population. However, bFGF affects the development of bipolar AChE-positive neurons, probably belonging to the non-motoneuron population. The data indicate that bFGF and aFGF are mitogens for neuroblasts from rat spinal cord in culture and that bFGF influences the development of a subpopulation of spinal neurons that are AChE-positive.