Zhu Feng, Nie Rong-Rong, Wu Ling, Liu Lei, Tang Wei, Tian Wei-Dong
State Key Laboratory of Oral Diseases, West China Stomatological Hospital, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2008 Mar;39(2):290-3.
To observe the overexpression of dentin sialophosphoprotein (DSPP) in mouse bone marrow mesenchymal stem cells (BM-MSC) and the expression of specific gene involved in odontogenic and osteogenic differentiation in transfected cells.
Mouse bone marrow mesenchymal stem cells (BM-MSC) were cultured and then transfected with DSPP gene by adenovirus-mediated way in vitro. The transfection efficiency of DSPP gene was assessed by the marker gene green fluorescent protein (GFP). With having observed the morphological transformation of transfected BM-MSC, we studied whether the transfected BM-MSC would express odontogenic and osteogenic genes by RT-PCR.
The mouse BM-MSC were obtained and the adenovirus mediated DSPP gene used to transfect BM-MSC successfully. The transfected efficiency was 42.7%. The transfected BM-MSC were induced to differentiate into odontoblast-like cells and meanwhile expressed specific odontogenic and osteogenic differentiation genes such as DSPP, DMP1, Msx1/2, Pax9, Lhx6/7, Sox9, Cbfa1, Osx, Col I.
The bone marrow mesenchymal stem cells can make differentiation with the overexpression of dentin sialophosphoprotein and express specific odontogenic and osteogenic differentiation genes.
观察牙本质涎磷蛋白(DSPP)在小鼠骨髓间充质干细胞(BM-MSC)中的过表达情况以及转染细胞中牙源性和成骨分化相关特定基因的表达。
培养小鼠骨髓间充质干细胞(BM-MSC),然后在体外通过腺病毒介导的方式用DSPP基因进行转染。通过标记基因绿色荧光蛋白(GFP)评估DSPP基因的转染效率。在观察转染的BM-MSC形态转化后,通过逆转录聚合酶链反应(RT-PCR)研究转染的BM-MSC是否会表达牙源性和成骨基因。
成功获得小鼠BM-MSC,并利用腺病毒介导的DSPP基因成功转染BM-MSC。转染效率为42.7%。转染的BM-MSC被诱导分化为成牙本质样细胞,同时表达特定的牙源性和成骨分化基因,如DSPP、DMP1、Msx1/2、Pax9、Lhx6/7、Sox9、Cbfa1、Osx、I型胶原(Col I)。
骨髓间充质干细胞可通过牙本质涎磷蛋白的过表达进行分化,并表达特定的牙源性和成骨分化基因。