Strong M J, Garruto R M
Laboratory of Central Nervous System Studies, National Institute of Neurological Disorders and Stroke, Bethesda, Maryland 20892.
In Vitro Cell Dev Biol. 1989 Oct;25(10):939-45. doi: 10.1007/BF02624007.
The spinal cords of fetal NIH:CR mice, gestational age Day 12 to 14, were dissected free of meninges and dorsal root ganglia, chemically dissociated, and layered onto discontinuous Percoll gradients at densities 1.040, 1.050, and 1.060 g/ml. After centrifugation (800 Xg for 15 min at 4 degrees C), three morphologically, biochemically, and immunohistologically distinct cell populations were collected from the gradient interfaces. The first interface, located at a density of 1.040 g/ml, was choline acetyltransferase enriched (0.86 +/- 0.08) compared to the second and third fractions (0.42 +/- 0.01 and 0 pmol acetylcholine synthesized/microgram protein, respectively). When simultaneously cultured with fetal mouse cardiac muscle on a gelatin-polylysine-laminin substrate in serum-free medium, these cells developed the characteristics of motor neurons.
妊娠12至14天的胎鼠NIH:CR品系小鼠的脊髓被解剖，去除脑膜和背根神经节，进行化学解离，并分层置于密度为1.040、1.050和1.060 g/ml的不连续 Percoll 梯度液上。离心（4℃下800×g离心15分钟）后，从梯度界面收集到三个在形态学、生物化学和免疫组织学上不同的细胞群体。第一个界面位于密度1.040 g/ml处，与第二和第三部分（分别为0.42±0.01和0 pmol乙酰胆碱合成/微克蛋白质）相比，富含胆碱乙酰转移酶（0.86±0.08）。当在无血清培养基中的明胶-聚赖氨酸-层粘连蛋白底物上与胎鼠心肌同时培养时，这些细胞呈现出运动神经元的特征。
