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

作　　者：马勇[1,2] 郭杨[1] 鲁俊山[1] 黄桂成[1] 王建伟[3] 

机构地区：[1]南京中医药大学骨伤研究所,江苏南京210023 [2]南京中医药大学附属医院骨伤科,江苏南京210029 [3]南京中医药大学无锡附属医院骨伤科,江苏无锡214001

出　　处：《中国骨质疏松杂志》2015年第1期13-18,共6页Chinese Journal of Osteoporosis

基　　金：江苏省中医药领军人才培养对象资助项目(LJ200924);江苏省自然基金资助项目(BK2011812)

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BMP-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〈0.05）,且5%中药＋BMP-2组优于BMP-2组（P=0.041）,增殖高峰出现在干预后第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BMP-2组及BMP-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。Objective To observe the effect of kidney-tonifying Chinese herb containing serum combined with BM P-2 on the morphology and proliferation of human umbilical cord blood mesenchymal stem cells（ UCB-M SCs）.Me thods The morphology change of primary,passaged,and intervened UCB-M SCs was observed using an inverted microscope.The third passage of UCBM SCs was randomly divided into 8 groups culturing in different concentrations of kidney-tonifying Chinese herb containing serum and BM P-2.The cell proliferation was detected using M TT method at 1-,3-,and 5-day after the intervention.Re sults The number of primary UCB-M SCs increased and the shape enlarged gradually along with the culturing time.After 2 weeks,most of the UCB-M SCs 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 BM P-2 group increased after 3-and 5-day intervention（ P〈0.05）.The effect in 5% combining group was superior to that in BM P-2 group（ P = 0.041）.The proliferation peak appeared on the third day after the intervention.The UCB-M SCs became cubical or polygonal shape in 3 days after the conditioned culture,especially in 5% Chinese herb containing serum combined with BM P-2 group and BM P-2 group.Conclusion Kidney-tonifying Chinese herb containing serum combined with BM P-2 can synergistically promote the proliferation of UCB-M SCs.The stimulation is time and concentration dependent.

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 

分 类 号：R274.9[医药卫生—中西医结合]