摘要
目的探讨p53抑制剂(PFT-α)与骨形态发生蛋白2(BMP-2)联合诱导对骨髓间充质干细胞(BMMSCs)向心肌样细胞分化的影响。方法分离大鼠BMMSCs进行培养、传代及纯化,分别应用PFT-α、BMP-2及两者联合对第2代BMMSCs定向诱导,不加诱导剂为对照组。倒置显微镜下观察并记录对数增殖期细胞的形态变化;流式细胞计数法鉴定BMMSCs表面联合标记物;免疫组化法检测c Tn I、Cx43及TPM的表达;免疫荧光检测技术对c Tn I和Cx43进行荧光标记;透射电镜观察分化细胞的超微结构变化。结果经诱导后的BMMSCs以梭形为主,细胞体积变大。CD45、CD29和CD90的阳性率分别为0.3%、86.5%和84.7%。诱导组细胞的c Tn I、Cx43及TPM均呈阳性表达,对照组为阴性。诱导组细胞胞质内Cx43免疫荧光染色表达呈红色,c Tn I则呈绿色,胞核椭圆位于细胞中央,胞质内可见平行排列的肌丝、线粒体和糖原等。结论 PFT-α和BMP-2均可单独诱导BMMSCs分化为心肌样细胞,两者联合应用更加高效、安全。
Objective To investigate the effect of PFT-α and BMP-2 on the differentiation of bone marrow mesenchymal stem cells（ BMMSCs） into heart like cells. Methods Cells were isolated from rat bone marrow and cultured,passaged and purified method,PTF-α,BMP-2 were used to induce the second generation BMMSCs,and the inducer was not used as the control group.The application of inverted microscope to observe the changes of growth and morphology of cells cultured by flow cytometry microscope,identified by BMMSCs combined with surface markers were detected by immunohistochemical method c Tn I,Cx43 and TPM expression and ultrastructural changes were observed in the differentiated cells by transmission electron microscope. Results After induction,BMMSCs was spindle shaped,the volume of cells became larger.CD29（ ＋） 86. 5%,CD45（-） 0. 3%,CD90（ ＋） 84. 7%.The expression of c Tn I,Cx43 and TPM in the induced group was positive.The expression of Cx43 in the cytoplasm of the induced group was red,and the expression of c Tn I immunofluorescence staining was green. oval nucleus is located in the central cell,Golgi apparatus,mitochondria,rough endoplasmic reticulum and ribosome subcellular structures located incytoplasm,parallel arrangement of filaments. Conclusions Both PFT-α and BMP-2 can induce BMMSCs to differentiate into cardiomyocytes,and the combination of these two methods is safe and efficient.
作者
陈晓依
王浩宇
吕洋
王海萍
孙微
李柔
CHEN Xiao-yi;WANG Hao-yu;LYU Yang;WANG Hai-ping;SUN Wei;LI Rou(Basic Medical College;Dept.of Histology and Embryology,Hebei North University,Zhangjiakou 075000,China)
出处
《基础医学与临床》
CSCD
2018年第8期1088-1093,共6页
Basic and Clinical Medicine
基金
河北省教育厅项目(Z2015057)
河北省自然科学基金(H2015017)
河北北方学院创新培育(CXRC1315)
关键词
骨髓间充质干细胞
PFT-Α
BMP-2
心肌细胞
bone marl＇ow mesenchymal stem cells
PFT-α
bone morphogenetic protein 2
cardiomyocytes
