摘要
目的:研究促红细胞生成素(erythropoietin,Epo)对大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMMSCs)增殖和分化的影响。方法:分离培养大鼠BMMSCs并进行传代;在成骨诱导条件下,以腺病毒为载体,分别携带Epo基因和EGFP基因作为实验组和阴性对照组,转染第3代BMMSCs,以加入相同体积无血清培养基作为空白对照组;通过MTT、RT-PCR、碱性磷酸酶(alkaline phosphatase,ALP)染色以及茜素红染色方法检测细胞的增殖和分化;采用配对t检验进行统计学分析。结果:阴性对照组与空白对照组相比没有差异;实验组与阴性对照组相比,细胞数量增加(P<0.05),ALP染色以及钙结节形成明显;实验组3 d时Runx2、Osx的表达及9 d时OCN的表达增加(P<0.05)。结论:Epo能促进BMMSCs的增殖及其向成骨细胞的分化。
Objective： To study the effects of erythropoietin（Epo） on the proliferation and differentiation of bone marrow mesenchy- mal stem cells（BMMSCs） in vitro. Methods： BMMSCs were isolated and cultured in vitro. Adenovirus vector AdCMV-EGFP was used for gene transfection. BMMSCs of the third passage were respectively transfected by AdCMV-EGCP-Epo（experimental group）, AdCMV-EGFP（negative control） and the same medium（blank control） followed by osteogenic induction. MTY assay, RT-PCR,al- kaline phosphatase（ALP） staining and alizarin red staining were used to test the proliferation and differentiation of the cells. Statisti- cal analysis was conducted by t test. Results： There was no difference between the negative control and the blank control Ad-Epo transfection increased the proliferation of BMMSCs（P 〈 0.05 ）, ALP expression（P 〈 0. O1 ） and mineralized nodule formation（ P 〈 O. 05）, increased the expression of runx2, Osx and OCN（P 〈0.05）. Conclusion： Epo can promote the proliferation and osteoblastic differentiation of BMMSCs.
出处
《实用口腔医学杂志》
CAS
CSCD
北大核心
2014年第1期10-14,共5页
Journal of Practical Stomatology
基金
国家自然科学基金面上项目(编号:81271111)
关键词
促红细胞生成素
骨髓间充质干细胞
腺病毒
转染
骨形成
Erythropoietin （Epo）
Bone marrow mesenchymal stem cells
Adenovirus
Transfection
Bone for-mation
