摘要
本实验目的在于研究小鼠胚胎脊髓提取液对损伤环境中的体外培养脊髓和背根节神经细胞的保护作用。神经细胞取自１２～１４ ｄ 胚胎小鼠脊髓和背根节，接种于２４ 孔板中。培养第５ ｄ 时在培养板中加入２５０ ｍ ｇ／Ｌ 脊髓提取液，对照组不加；培养第１０ ｄ 在液体表面加入液体石蜡造成缺气损伤。继续培养４、８、１２、２４ ｈ，然后去除液体石蜡，观察细胞形态和Ｋ＋ 、ＬＤＨ 的变化，用Ｍ ＴＴ 方法检测细胞活性，通过ＮＳＥ免疫组化反应显示神经元存活状况。结果表明：损伤神经元出现细胞肿胀、失去折光性、核偏位以及ＮＳＥ 染色变弱；而在培养液中加入脊髓提取液，可以减轻这种“缺气”造成的损伤，提高体外培养神经元在损伤环境中的生存率，保持细胞的活性和细胞膜的通透性。
This study was undertaken to determine the effect of spinal cord crude extract(SCCE) of mouse embryo on cultured spinal cord and DRG neurons in damaged environment. Primary cultured neurons were dissociated from 12～14 day old mouse embryo and plated on 24 well cultureplates. During 5 DIV, 250 mg/L SCCE was added into the medium for experimental groups, and control group without SCCE. At 10 DIV, liquid paraffin was added carefully on the surface of medium to make an injurious condition and cultured for 4, 8, 12, 24 hrs. Then liquid paraffin was discarded off, finally we observed the morphology of cells and test levels of K +, LDH in this media. At the same time MTT was used to test cell activity, and immunocytochemistry of NSE was used to show the survival neurons. The results indicated that damaged neurons showed various changes such as cell swelling, lacking of dioptre, nucleus shifting to the cell periphery, weaker NSE staining and so on, after being sealed with paraffin oil for different times. But the present results with SCCE in this study demonstrated that SCCE could reduce these kinds of gas deprivated injury and improved neuronal survival rate in vitro under the damage environment as well as maintaining cell activity and permeability of cell membrane. (Figures 3,4 on plate 20,21)
出处
《神经解剖学杂志》
CAS
CSCD
北大核心
1999年第2期127-131,共5页
Chinese Journal of Neuroanatomy
关键词
脊髓提取液
神经细胞
细胞培养
缺气损伤
spinal cord extract, neuron, cell culture, gas deprivated injury, rat
