Development of dual promoter-driven reporter systems and their application in simultaneous tracing of mesenchymal stem cells differentiating into the cardiomyogenic or the endothelial cell lineage

摘要

A recombinant expression vector for monitoring the differentiation of a cardiomyogenic cell and endothelial cell is provided to detect the cell differentiation after cell therapy and verify the stability. A recombinant expression vector pMLC-2v-DsRed-Flk-1-EGFP or pMLC-2v-DsRed-Tie-2-EGFP for monitoring the differentiation of a cardiomyogenic cell and endothelial cell comprises a promoter which induces the cardiomyogenic cell-specific expression; a first reporter gene which is connected to be operatable; a promoter which induces the endothelial cell-specific expression; and a second reporter gene which is connected to be operatable. The promoter which induces cardiomyogenic cell-specific expression is the myosin light chain 2v promoter of the sequence number 1(SEQ ID NO:1), rat myosin light chain 2v promoter of the sequence number 2, or myocardium troponin I promoter of the sequence number 3. The endothelial cell-specific promoter is the Flk-1 promoter of the sequence number 4 and Tie-2 promoter of the sequence number 5. A method for monitoring the differentiation of cardiomyogenic cell and endothelial cell at once: a step of preparing a mesenchymal stem cell; a step of inducing the recombinant expression vector for monitoring the differentiation of cardiomyogenic cell and endothelial cell into the mesenchymal stem cell; and a step of confirming the expression of reporter gene in a transformed mesenchymal stem cell.