人骨髓间充质干细胞在体外向成骨细胞定向分化的研究分析  

The Basic Research Analysis of Direct Differentiation of Human Bone Marrow Mesenchymal Stem Cells into Osteoblasts in Vitro

作　　者：刘志刚[1] 熊敏[1] 唐冰[1] 李锋[2] 余化龙[1] 曾云[1] 陈洁[1] 何宁[1] 王志勇[1] 韩珩[1] 

机构地区：[1]湖北医药学院附属东风医院骨科研究所,湖北十堰442000 [2]华中科技大学同济医学院附属同济医院骨科,武汉430030

出　　处：《医学综述》2015年第18期3412-3414,共3页Medical Recapitulate

基　　金：十堰市科学技术研究与开发项目计划(14Y50)

摘　　要：目的探讨人骨髓间充质干细胞在体外向成骨细胞定向分化情况。方法取引产胎儿的人骨髓间充质干细胞进行原代细胞培养,诱导培养向成骨细胞分化,诱导后进行形态学观察,碱性磷酸酶(ALP)染色与生长曲线测定。当人骨髓间充质干细胞培养生长密度为80.0%左右时分为转染组与对照组,对照组不进行转染,转染组常规进行转染。结果原代细胞生长2 h开始贴壁,培养4 d左右胞体逐渐变得粗大。经成骨诱导后细胞形态由长梭形逐渐变为多边形,形成细胞结节甚至团块状。转染组的胎儿骨髓间质干细胞和未转染对照组的生长状况对比差异无统计学意义(P>0.05),转染细胞与未转染细胞倍增时间分别为(23±4)h和(25±4)h。转染组在转染后第3、6、9、12日的ALP活性均明显高于对照组[(0.12±0.02)%比(0.10±0.02)%,(0.16±0.02)%比(0.12±0.02)%,(0.20±0.03)%比(0.13±0.02)%,(0.23±0.04)%比(0.14±0.02)%],差异有统计学意义(P<0.01)。结论胎儿骨髓间充质干细胞在成骨细胞分化转染的诱导下可促进向成骨细胞进行定向分化,为胎儿间质干细胞分化调控和药物作用机制研究奠定基础。Objective To discuss human bone marrow mesenchymal stem cells differentiation into osteoblasts in vitro. Methods Human bone marrow mesenchymal stem cells of induced labor fetus were taken for primary culture to induce osteoblast differentiation,the morphology was observed,the alkaline phosphatase（ ALP） staining and the growth curve were determined. When human bone marrow mesenchymal stem cell culture growth density was about 80. 0%,they were divided into two groups： transfection group and control group,the control group was not transfected,the transfection group was givenconventional transfection.Results The primary cell were adherent growth after 2 h and showed stretch fusiform adherent cells after cultured for 4 days. The cell body gradually became thick and showed elongated protrusions. After inducing,the cells showed morphology. The cells growth conditions between the transfected fetal bone marrow mesenchymal stem cells and the untransfected cells showed no significant difference（ P 〉0. 05）. The doubling time of the transfected cells and non-transfected cells were（ 23 ± 4） h and（ 25 ± 4） h respectively. The ALP activity of the transfection group was significantly higher at different times [（ 0. 12 ± 0. 02） % vs（ 0. 10 ±0. 02） %,（ 0. 16 ± 0. 02） % vs（ 0. 12 ± 0. 02） %,（ 0. 20 ± 0. 03） % vs（ 0. 13 ± 0. 02） %,（ 0. 23 ±0. 04） % vs（ 0. 14 ± 0. 02） % ]（ P〈0. 01）. Conclusion Human bone marrow mesenchymal stem cells directly differentiated into osteoblasts in vitro under induction,which can be the basis for further fetal interstitial stem cell differentiation regulation and study on the drug action mechanisms.

关 键 词：人骨髓间充质干细胞 成骨细胞 定向分化 地塞米松 碱性磷酸酶 

分 类 号：R468.2[医药卫生—临床医学]