Dissociated dorsal root ganglia in tissue culture.
Dissociated dorsal root ganglia in tissue culture.
复制标题
组织培养中分离的背根神经节。
DOI：
10.1002/aja.1000990105
复制
发表时间：
1956
期刊：
影响因子：
--
通讯作者：
J. Nakai
中科院分区：
文献类型：
--
作者：
J. Nakai
材料和方法本研究所用脊神经节取自8日龄至12日龄鸡胚胎的腰、骶部和胸区。图5和图6说明了典型材料的大小、位置和关系。对年轻动物的解剖被发现是相当繁琐的，特别是在分离脊神经节和自主神经链方面。分离是通过在Kahn管中转移20到50个神经节和1ml含有500到2500个单位的蛋白酶A4的盖氏平衡盐溶液(BSS)并在37℃下孵育15到60分钟来完成的。然后用BSS以温和的吸管作用洗涤组织，然后在600rpm下离心5分钟。这一过程重复了两到三次。将一滴沉淀物加入等量的8日龄雏鸡胚胎提取液中，或与人胎盘血清和牛血清超滤液等量联合使用。加入一滴肝素化的公鸡血浆产生凝血。从20到50个神经节的游离元素被用来建立大约24个Maximow玻片制剂。研究发现，添加来自同一胚胎的椎骨、胸壁或坐骨神经等组织片段有利于神经纤维的生长。
MATERIALS AND METHODSThe spinal ganglia employed in the present study were taken from the lumbo-sacral and thoracic regions of 8-to 12-day chick embryos. Figures 5 and 6 illustrate the size, location and relations of typical material. Dissection of younger animals was found to be considerably more tedious, especially in separating spinal ganglia from the autonomic chain.Dissociation was accomplished by transferring 20 to 50 ganglia in Kahn tubes with 1ml of Gey’s balanced salt so-lution (BSS) containing from 500 to 2500 units of proteinase A 4 and incubated at 37 C. for 15 to 60 minutes. The tissues were then washed with BSS using gentle pipette action and then centrifuged at 600rpm for 5 minutes. This procedure was repeated two or three times. One drop of the sediment was added to an equal amount of embryonic extract derived from 8-day chicks alone or in equal combination with human placental serum and ox serum ultrafiltrate. Clotting was produced with the addition of one drop of heparinized cockerel plasma. The dissociated elements from 20 to 50 ganglia were used to set up approximately 24 Maximow slide preparations. The addition of tissue fragments such as vertebrae, chest wall or the sciatic nerve from the same embryo was found to favor the outgrowth of nerve fibers.