色谱2016，Vol.34Issue(4)：414-421,8.DOI:10.3724/SP.J.1123.2016.01006
多层面验证共培养微环境诱导法诱导骨髓间充质干细胞心肌样分化
Feasibility of cardiac differentiation of bone marrow mesenchymal stem cells induced by co-culture indirectly with cardiomyocytes in several dimensions
摘要
Abstract
The objective of this research is to investigate the feasibility of cardiac differentiation of bone marrow mesenchymal stem cells( MSCs)by co-culture with cardiomyocytes( CMs)in vitro. The third generation of MSCs from bone marrow and CMs were co-cultured indirectly in a transwell. One week later,the expressions of muscle-specific markers( cardiac troponin I andα-actin) by immunofluorescence staining and the gene expressions of transcription factors ( Nkx-2. 5 and GATA-4 ) were measured by real-time polymerase chain reaction ( RT-PCR ). Then,orthogonal partial least squares discriminant analysis( OPLS-DA)models were employed to confirm the difference among MSCs,induced MSCs and CMs by ultra-performance liquid chromatography-tandem mass spectrometry( UPLC-MS/MS)analysis. The distinctive changes were identified by multivariate analysis and variate analysis,and the changed metabolites were identified by MS and MS/MS. One week after co-cultured with CMs indirectly,the specific myo-cardium morphology of MSCs was observed under microscope. The positive immunofluores-cence stainings against cTnI and α-actin were detected,and the positive expressions of the transcription factors Nkx-2. 5 and GATA-4 were measured by RT-PCR. In OPLS-DA mode,obvi-ous trend of cardiac differentiation of MSCs can be seen,and seven metabolites were tested both in induced MSCs and CMs,but five metabolites were tested in induced MSCs or CMs. Car-diac differentiation of MSCs can be induced by co-cultured with CMs indirectly in vitro. However,metabolism difference still existed between induced MSCs and CMs.关键词
共培养微环境/代谢组学/心肌样分化/骨髓间充质干细胞/心肌细胞Key words
co-culture microenvironment/metabonomics/myocardium-like differentiation/bone marrow mesenchymal stem cells( MSCs)/cardiomyocytes( CMs)分类
化学化工引用本文复制引用
何林静,邓伏雪,胡云凤,朱卫文,伍家燕,常静..多层面验证共培养微环境诱导法诱导骨髓间充质干细胞心肌样分化[J].色谱,2016,34(4):414-421,8.基金项目
国家临床重点专科建设项目（2011170）．@@@@National Key Clinical Specialties Construction Program of China（ No．2011170）． （）
