摘要
目的:应用神经生长因子培养胎兔许旺细胞,观察许旺细胞生长的情况。方法:实验于2002-02/2004-05在河北医科大学第三临床医学院中心实验室和细胞培养室完成。怀孕28d的新西兰白兔,剖腹取出胎兔,取胎兔坐骨神经,剥干净外膜,用植块加差速贴壁联合法培养胎兔许旺细胞,纯化传代后的胎兔许旺细胞分成2组培养,一组加含神经生长因子的培养液,即实验组,另一组加常规培养液,即对照组。细胞传代后24h在每个培养皿中随机选取3个视野,开始初次细胞计数,以后在统一视野每天进行许旺细胞计数。增殖指数=各次观察细胞数/初次观察细胞数。结果:实验用剖腹取出胎兔8只,坐骨神经16根。60个观察视野,均进入结果分析。①在第6天见细胞从胎兔神经组织块周围爬出。②第8天细胞沿组织块向外生长,可见大量梭形的胎兔许旺细胞;第14天植块周围细胞长势最好。③有两种细胞,一种为细胞形态梭形,有突起,多为双极,偶尔可见3个细长突起,胞核呈椭圆形,胞体周围有光晕,即生长晕,为许旺细胞。偶尔可见另一种细胞,胞体大,呈扁平状,外形不规则,突起短而多,细胞核较浅,核仁明显,有2～3个核仁,为成纤维细胞。④根据许旺细胞特有的特征性形态以及细胞免疫组化染色可证实为许旺细胞。⑤实验组许旺细胞的分裂增殖速度明显比对照组加快犤传代培养后第3天:(1.41±0.12),(1.05±0.10);第4天:(1.85±0.26),(1.21±0.22);第5天:(2.36±0.43),(1.23±0.32);第6天:(3.52±0.11),(1.94±0.33);第7天:(6.25±0.38),(3.14±0.27),P<0.001犦。结论:用神经生长因子可促进胎兔许旺细胞生长分裂,能够获得大量的胎兔许旺细胞,满足组织工程对许旺细胞的需求;此外胎兔许旺细胞作为组织工程的种子细胞也可以减轻免疫排斥反应的发生。
AIM： To observe the growth of Schwann cells of fetal rabbits by applying nerve growth factor （NGF） in culture. METHODS： The experiment was conducted at the Central Laboratory and Laboratory of Cell Culture, Third Clinical Medical College of Hebei Medical University from February 2002 to May 2004. Totally 28 pregnanry New Zealand white rabbits were paunched . Fetal rabbits were taken out from their abdomens rapidly. Then the sciatic nerve of fetal rabbits was fetched and its ectoblastic was removed. We adopted a method of transplant piece and speed difference adhesion to cultivate Schwann cells. The purified and generative Schwann cells were divided into 2 groups for culture： experimental group （Culture medium of nerve growth factor was added） and control group （Routine culture medium was added）. Twenty-four hours after cell passage, 3 visual fields were randomly chosen from each Petri dish. Cell count of Schwann cells began for the first time, then conducted it at the same field every day. Proliferative index=Cell number each time/Cell number at the first time. RESULTS： There were 8 fetal rabbits, 16 sciatic nerves and 60 visual fields used in the experiment; all of them entered the stage of result analysis.① On the 6^th day, Cells were seen around the fetus rabbit sciatic piece in the utensil. ② On the 8^th day, a lot of Schwann cells presenting shuttle were seen around the fetus rabbit sciatic piece in the Petri dish. Schwann cells grew best on the 14^th day.③ There were two types of cells. One was shuttle morphology with 3 slightness prominence seen occasion- ally and a majority of cells had double poles and ellipse nuclear. There was a light circle named growth faint in the vital cell. This was Schwann cell. Occasionally, another cell was visible in the utensil. Its body was big and flat. The morphology was unregular and there were many short prominences and the light nuclear of cell had 2-3 obvious nucleoluses. This was fibroblast. ④ The Schwann cells were confirmed according to r
出处
《中国临床康复》
CSCD
北大核心
2006年第9期63-65,共3页
Chinese Journal of Clinical Rehabilitation
基金
河北省教委资助项目(2002234)~~
