摘要
目的探讨脊髓前角运动神经元在间接共培养情况下是否可以对雪旺细胞(Schwann cells,SCs)的功能产生影响。方法从孕15d的SD胎鼠的脊髓内分离培养脊髓前角运动神经元;从新生1d的SD大鼠的坐骨神经内分离培养SCs。将上述两种细胞分别种植于Transwell培养板的上、下室内,通过细胞计数、3H-TdR掺入观察SCs的存活与增殖情况,Western blot检测SCs表达脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)情况。然后将经共培养处理和未经共培养处理的SCs分别与经1%低氧预处理4h的脊髓前角运动神经元共培养72h,通过细胞计数、MTT法检测脊髓前角运动神经元存活情况。结果共培养组SCs增殖指数为(24197.6±739.8),明显优于直接培养组的(17451.2±512.7)(P<0.05),共培养组SCs表达BDNF的水平明显高于直接培养组(P<0.05)。缺氧后处理组脊髓前角运动神经元存活数量及MTT分别为(89.6±2.3)、(0.2384±0.0067),分别优于对照组的(66.4±4.8)、(0.1962±0.0059)(P<0.05)。结论在间接共培养情况下,脊髓前角运动神经元可以促进SCs的增殖及分泌BDNF等功能;SCs对缺氧后的脊髓前角运动神经元具有更明显的保护作用。利用Transwell进行间接共培养,可以获得活化状态的SCs。
Objective To investigate the interaction between motoneurons （MNs） and Schwann cells （SCs） in co-culture in Transwell system. Methods MNs were harvested from SD rats at embryonic day 15; SCs were obtained from sciatic nerve of neonatal rats at day 1. MNs were cultured in the upper room of Transwell, while SCs in lower room （SCs pretreatment）. The proliferation of SCs was detected by cell count and ^3H- TdR incorporation, and the level of BDNF in SCs was detected by Western blot. The SCs pretreated or not and MNs pre-cultured in 1% 02 for 4 h were co-cultured for 72 h, then the survival amount of MNs was detected by cell count and MTI＇. Results The proliferation index of SCs was （24 197.6 ± 739.8） after co-culture with MNs in Transwell （group A） and （17 451.2 ±512.7） in its control group （group B） （P 〈0.05）, and the level of BDNF in group A was higher than that in group B. The amount of hypoxic MNs and the value from MTI＂ were （89.6 _±2. 3）, （0.238 4 ±0.006 7） respectively after co-culture with pretreated SCs （ group C）, and （66.4 ± 4.8 ）, （0. 196 2 ± 0.005 9） in its corresponding control group （group D） （P 〈 0.05 ）. Conclusion MNs can enhance the survival of SCs and their BDNF expression in non-contact culture manner, and these SCs have obvious protective effect on hypoxic MNs.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2007年第23期2226-2229,共4页
Journal of Third Military Medical University
基金
国家自然科学基金(30400454)
国家重点基础研究发展规划项目("973"项目)(2005CB522604)~~
关键词
运动神经元
雪旺细胞
增殖
共培养
motoneuron
Schwann cell
proliferation
co-culture
