Experimental Study On LIM Mineralization Protein-1 Promote The Osteogenic Potential Of Cultured Bone Marrow Mesenchymal Stem Cells

Objective: Construct an eukaryotic expression plasmid carrying hLMP-1, and have it expressed in rabbit BMSCs, to study the influence of hLMP-1 on osteogenic potential of cultured BMSCs.Methods: Reverse transcription- polymerase chain reation (RT-PCR) technique, was used to clone hLMP-1 and hBMP-2 from human placenta tissue and plasmid with hBMP-2. Digestion snd ligations with eukaryotic expression plasmid pIRES2-EGFP, pIRES2-EGFP-LMP1 and pIRES2-EGFP-BMP2 recombinants were constructed. BMSCs were isolated and culture-expanded from rabbit bone marrow. The third culture was chosen for study. With the help of lipofectamine, plasmids pIRES2-EGFP-LMP1, pIRES2-EGFP-BMP2 and pIRES2-EGFP were transfected into BMSCs. Transfective rate was estimated under fluorescence microscope and cells was tested seven days later. The expression of LMP-1,BMP-2 and collagen typeâ… was tested with RT-PCR. ALP testing kit was used to measure ALP activity, and immunohistochemical staining was used to test the expression of osteocalcin.Results: Restriction and sequencing analyses confirmed that eukaryotic expression plasmid pIRES2-EGFP-LMP1 and pIRES2-EGFP-BMP2 was...