电磁场不同处理时间对人脐带间充质干细胞增殖与分化的影响  

Effect of Electromagnetic Field in Different Treatment Time on Proliferation and Differentiation of Human Umbilical Cord of Mesenchyme Stem Cells in Vitro

作　　者：周建[1] 马小妮[1] 陈克明[1] 闫娟丽[1] 高玉海[1] 石文贵[1] 方清清 谢艳芳[1] 

机构地区：[1]兰州军区兰州总医院骨科研究所,兰州730050

出　　处：《解放军医药杂志》2015年第3期11-15,19,共6页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：国家自然科技基金(81270963;81471090);甘肃省科技重大专项资助项目(09ZNKDA025)

摘　　要：目的研究频率50 Hz强度1.8 m T下的不同处理时间正弦交变电磁场对体外培养人脐带间充质干细胞增殖与分化的影响。方法体外分离培养人脐带间充质干细胞,传代后随机分为6组,每天于倒置相差显微镜下观察细胞形态;MTT测定细胞增殖;10 d、12 d、14 d和16 d测定ALP活性;15 d行茜素红钙化结节染色以及17 d行vonkossa染色;第13 d ALP组织化学染色;在磁场处理后的4 d和6 d行Real-time RT-PCR检测Ⅰ型胶原(Collagen-Ⅰ)和骨形态发生蛋白(BMP-2)mRNA表达量的变化。结果 0.5 h组可促进细胞增殖;磁场处理10 d^12 d后细胞出现钙化趋势;2.5 h组在SEMFs处理16 d和18 d后ALP活性均高于对照组(P<0.05);1.5 h组、2.0 h组和2.5 h组钙化结节茜素红染色面积均高于对照组(P<0.01,P<0.05);1.5 h组Collagen-ⅠmRNA表达水平高于对照组(P<0.05),2.0 h和2.5 h组ALP组织化学染色面积、Collagen-Ⅰ、BMP-2 mRNA表达水平均高于对照组(P<0.05,P<0.01)。结论 50 Hz 1.8 m T的正弦交变磁场能促进体外培养人脐带间充质干细胞成骨性分化,尤以处理2.5 h促进人脐带间充质干细胞成骨性分化最为明显。Objective To investigate effect of different treatment time of sinusoidal electromagnetic fields（ SEMFs） under 50 Hz 1. 8 m T on proliferation and differentiation of human umbilical cord of mesenchyme stem cells in vitro. Methods The human umbilical cord of mesenchyme stem cells were isolated in vitro,and randomly divided into6 groups after one passage. The cell configurations were observed every day under inverted phase contrast microscope;MTT colorimetric assay was used to detect the cell proliferation; ALP activities were detected on 10 thd,12thd,14 thd and16thd after SEMFs treatment; calcified nodules were stained with alizarin red on 15 thd and with vonkossa staining on17 thd after SEMFs treatment; ALP histochemical staining was performed on 13 thd after SEMFs treatment; the expression changes of Collagen-Ⅰ and BMP-2 mRNA were detected with real-time RT-PCR（ reverse transcription polymerase chain reaction） on 4thd and 6thd after SEMFs treatment. Results The SEMFs promoted the cell proliferation in 0. 5 h group;the calcified tendency was found on 10thd-12 thd after SEMFs treatment; the ALP activities on 16 thd and 18 thd after SEMFs treatment in 2. 5 h group were significantly higher than those in control group（ P〈0. 05）; the areas of alizarin red of calcified nodules in 1. 5 h,2. 0 h and 2. 5 h groups were significantly larger than that in control group（ P〈0. 01,P 0. 05）; Collagen-Ⅰ mRNA expression level in 1. 5 h group was higher than that in control group（ P〈0. 05）,and ALP histochemical staining areas,expression levels of Collagen-Ⅰ and BMP-2 mRNA in 2. 0 h and 2. 5 h groups were significantly larger and higher than those in control group（ P〈0. 05,P 0. 01）. Conclusion Sinusoidal electromagnetic fields under 50 Hz 1. 8 m T can promote osteoplastic differentiation of human umbilical cord of mesenchyme stem cells in vitro,and the differentiation is the most obvious in 2. 5 h group.

关 键 词：间质干细胞 脐带 电磁场 细胞分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]