骨髓基质细胞条件液对PC12细胞生长与分化的影响  

Effects of marrow stromal cells conditioned medium on proliferation and differentiation of PC12 cells

作　　者：顾平[1] 刘力[1] 王铭维[2] 王彦永[2] 崔冬生[1] 

机构地区：[1]河北省脑老化与认知神经科学实验室,河北省石家庄市050031 [2]河北医科大学第一医院神经内科,河北省石家庄市050031

出　　处：《中国临床康复》2006年第33期16-18,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金资助项目(30400138);河北省自然科学基金资助项目(C2004000583)~~

摘　　要：目的:观察骨髓基质细胞条件液对PC12细胞生长与分化的影响,分析其能否保护PC12细胞免受鱼藤酮的毒性作用。方法:实验于2005-11/2006-04在河北省脑老化与认知神经科学重点实验室完成,PC12细胞来源于大鼠嗜铬细胞瘤。①条件液对PC12细胞增殖影响和对毒物鱼藤酮的抵抗作用:PC12细胞按实验对照原则分为对照组、条件液组,分别经正常培养液和骨髓基质细胞培养制备的条件液培养。两组分别加入鱼藤酮,浓度为0和10μmol/L,作用72h,MTT法检测细胞活性。细胞存活率=鱼藤酮孔吸光度值/对照孔吸光度值×100%。②条件液对PC12细胞的诱导分化作用:分别于接种第3,7,10天于倒置显微镜下观察细胞分化情况和突起生长情况。计数10～20个视野,按突起细胞数、3个或3个以上突起细胞数、突起长度大于2倍胞体细胞数占细胞总数的百分率3个指标对条件液组和对照组进行比较。结果:①条件液对PC12细胞增殖的影响及对毒物鱼藤酮的抵抗作用:经鱼藤酮作用后条件液组细胞存活率显著低于对照组[(64.45±1.60)%,(78.86±4.95)%(t=6.783,P<0.01)]。用条件液培养PC12细胞72h,条件液组与对照组的细胞存活率相比差异无显著性意义[(118.1±3.28)%,(121.3±10.54)%(P>0.05)]。②条件液对PC12细胞的诱导分化作用:条件液作用3d,细胞开始长出突起,但与对照组无差别。至第7天,有突起细胞数、突起大于2倍胞体直径的细胞数及有3个以上突起的细胞数均增多,占总细胞数的百分率与对照组比较差异均具有显著性意义(P<0.05～0.01)。培养至第10天,长度大于2倍胞体直径的细胞数占总细胞数的百分率与对照组比较差异也有显著性意义(P<0.01)。结论:骨髓基质细胞条件液可诱导PC12细胞分化为神经元样细胞,但对其增殖及鱼藤酮所致氧化损伤均无明显作用。骨髓基质细胞的这种诱导作用可能由其分泌到细胞外的可溶性分子介导AIM： To study the effects of marrow stromal cells conditioned medium （CM） on proliferation and differentiation of PC 12 cells, meanwhile whether CM can protect PC12 cell from the damage of rotenone. METHODS： The experiment had been done in the Hebei Provincial Brain Ageing and Cognitive Neuroscience Key Laboratory from November 2005 to April 2006. PC12 cells came from rat chromaffin tumor. ①Effect of CM on the proliferation, of PC 12 cell and the resistance to rotenone： PC 12 cells were divided into control group and CM group according to control principle, that were the normal medium and marrow stromal cell conditioned medium. The two group were added to rotenone, and the concentrations were 0 and 10 μmol/L. After cultured for 72 hours, the cell viability of PC12 was determined by MTr chromatometry. The survival rate was equal to the absorbance （A） of rote.none/the A of control group ×100%. ②The differentiation of PC12 cell with conditioned medium： The cell differentiation and apophysis growth of PC12 cell were observed under the invert microscope at days 3, 7 and 10. About 10 to 20 visual fields counted and the conditioned medium group and control group were compared according to the cell counts of apophysis and twice of diameter and three or more than three apophysis. RESULTS： ①Effect of conditioned medium on the proliferation of PC12 cell and the resistance to rotenone：＂ After cultured in rotenone, the survival rate of conditioned medium group was markedly lower than that of control group [（64.45±1.60）%, （78.86±4.95）% （t=6.783,P 〈 0.01）]. After euhured for 72 hours, there was no significant difference in survival rate between conditioned medium group and control group [（ 118.1±3.28）%, （121.3±10.54）% （P 〉 0.05）]. ②the differentiation of PC12 cell with eonditioned medium： The cell began to grow apophysis, but had no significant difference with control group after euhured in conditioned medium for 3 days. Until the seventh day the cell counts

关 键 词：骨髓细胞 PC12细胞 细胞分化 

分 类 号：R394.2[医药卫生—医学遗传学]