Expression and mechanism of cDNA of human bone morphogenetic protein 2 via eukaryotic expression vector in rabbit mesenchymal stem cells

Objective To investigate the transcription and expression of hBMP- 2 cDNA via eukaryotic expression vector in rabbit MSCs in vitro so as to provide materials for further experiments on treatment of bone defects by bionic nanobone carried with multigene- transfected MSCs. Methods The pIRES2- EGFP- hBMP2 plasmids were transfected into rabbit MSCs by means of electroporation. The efficiency of transfection was evaluated by fluorescence microscopy and flow cytometry assay. Quantitative RT- PCR was resorted to for observation of successful transcription of hBMP- 2 gene. Immunohistochemistry and western blot assay were used to identify the protein expression of hBMP- 2 gene. The function of hBMP- 2 was showed by quantitative analysis of ALP activity. Results Plasmids pIRES2- EGFP- hBMP2 could be transfected into rabbit MSCs through electroporation method. Green fluorescent protein was visible under fluorescence microscopy and the transfection efficiency was (42.7± 2.1)% by flow cytometry assay. The hBMP- 2 gene fragments were detected by RT- PCR. The protein was observed to be strongly positive in immunohistochemistry and had 18 KD demonstration in western blot assay. The ALP activity of tranfected MSCs was obviously increased( P 0.05). Conclusion The recombinant plasmid containing hBMP- 2 cDNA can be transfected and expressed effectively in rabbit MSCs