补肾中药血清联合BMP-2对人脐血间充质干细胞 增殖及形态学的影响
Effect of the combination of kidney-tonifying Chinese herb containing serum and BMP-2 on the proliferation and morphology of human mesenchymal stem cells derived from umbilical cord blood
  
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中文关键词:  中医中药  补肾中药  BMP-2 ;脐血间充质干细胞  细胞增殖
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马勇 郭杨〃鲁俊山1黄桂成1王建伟3 南京中医药大学骨伤研究所,江苏南京210023 
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中文摘要:
      目的观察补肾中药血清联合BMP-2对人脐血间充质干细胞(UCB-MSCs)增殖及形态学的影响,并探讨其最佳时效。 方法倒置显微镜观察原代、传代培养及干预后UCB-MSCs的形态学变化;取第3代UCB-MSCs,随机分为8组,给予不同浓度 补肾中药血清及腿P-2条件培养基培养,分别于诱导后1 d、3 d、5 d采用MTT法检测各组细胞增殖情况。结果原代培养 UCB-MSCs随时间延长,数量逐渐增多且逐渐变长,2周时多呈长梭形、旋涡状生长,传代培养3 d即达到70% ~ 80%融合,以 梭形为主,符合干细胞的生长特征;干预后第3、5 d,5%、10%中药+ BMP-2组和BMP-2组均能够促进UCB-MSCs增值(P <05),且5%中药+ BMP-2组优于BMP-2组(P = 0. 041),增殖髙峰出现在干预后第3 d;经条件培养3 d后,各组细胞均可见 由长梭形变成立方形或多角形,尤以5%中药联合腿P-2组及腿P-2组较为明显。结论补肾中药血清联合BMP-2可协同 促进体外培养UCB-MSCs增殖,增殖效应与干预时间和中药血清浓度有一定相关性。
英文摘要:
      Objective To observe the effect of kidney-tonifying Chinese herb containing serum combined with BMP-2 on the morphology and proliferation of human umbilical cord blood mesenchymal stem cells (UCB-MSCs). Methods The morphology change of primary’ passaged,and intervened UCB-MSCs was observed using an inverted microscope. The third passage of UCB- MSCs was randomly divided into 8 groups culturing in different concentrations of kidney-tonifying Chinese herb containing serum and BMP-2. The cell proliferation was detected using MTT method at 1-,3-,and 5-day after the intervention. Results The number of primary UCB-MSCs increased and the shape enlarged gradually along with the culturing time. After 2 weeks, most of the UCB-MSCs were characterized by spiral shape growth. In 3 days after passage’ 70% - 80% of the cells fused in the culture’ which met the growth characteristics of the stem cells. The cell proliferation in 5% and 10% combining groups and BMP-2 group increased after 3-and 5-day intervention (P <0. 05). The effect in 5% combining group was superior to that in BMP-2 group (P =0. 041). The proliferation peak appeared on the third day after the intervention. The UCB-MSCs became cubical or polygonal shape in 3 days
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