The Influence Of BMP₅ And BMP₉ In Myocardiocytes Differentiation Of Stem Cell In Vitro

PARTâ… THE CULTURE AND INDUCTION OF C₃H10T_1/2 STEM CELLS, THE CULTURE OF 293 CELLS, THE AMPLIFICATION OF THE ADBMP₅ AD-GFP AND ADBMP₉ AD-GFPObjective:Exploit the technique of cell inducement which can make the induction of C₃H10T_1/2 stem cell with AdBMP₅ and AdBMP 9 Ad-GFP , and amplifying the AdBMP₅ and AdBMP 9 Ad-GFP with 293 cells.The work above establishes the foundation of influence of BMP₅ and BMP 9 in myocardiocytes differentiation of stem cell In vitro.Methods :Amplify the AdBMP₅ and AdBMP 9 Ad-GFP with 293 cells. Infect the C₃H10T_1/2 stem cell with AdBMP₅ and AdBMP₉ Ad-GFP.Result:Amplify the AdBMP₅ and AdBMP 9 Ad-GFP with 293 cells successfully .The Infection efficiency of tansfecting the C₃H10T_1/2 stem cell with high titre AdBMP₅ and AdBMP₉ Ad-GFP is 36% and 34%.Conclusion:The titre of AdBMP₅ and AdBMP 9 Ad-GFP is improved by amplifying the AdBMP₅ and AdBMP 9 Ad-GFP with 293 cells. Infect ion of the C₃H10T_1/2 stem cell with AdBMP₅ and AdBMP₉ Ad-GFP successfully. PARTâ…¡THE INFLUENCE OF BMP₅ AND BMP 9 IN MYOCARDIOCYTES DIFFERENTIATION OF STEM CELL IN VITROObjective: Explore the influence of BMP₅ and BMP₉ in myocardiocytes differentiation of stem cell in vitro.Methods: The C₃H10T_1/2 stem cell were induced by 3.75 ml/L Ad-BMP₅ Ad-GFP and 2.67 ml/L Ad-BMP₉ Ad-GFP for 1 week and 2 weeks . The specific transcription factors of cardiomyocytes were detected in stem cells during differentiation with plasmid transfection by RT-PCR and the specific proteins in cardiomyocytes were detected by laser confocal microsopy After 1 week and 2 weeks;The change of cells ultramicro structure was detected by electron microscope after 2 weeks.Result:After induction for 1 week and 2 weeks : the volume of C₃H10T_1/2 stem cellsvolume are increscent obviously , the trend of cells become unanimous,the connection between cells are compact,refractivity of cells enhance conspicuously ;the expression of NKx2.5, GATA-4, MEF2C could be detected in C₃H10T_1/2 stem cells;Cx43 and cTnT can be detected with AdBMP₅ and AdBMP₉ Ad-GFP. The ultramicro structure Cardiomyocytes ultramicro structure can be detected after induction for 2 weeks.Conclusion:BMP₅ and BMP₉ make a very important influence in C₃H10T_1/2 stem cells targeted differentiation into cardiomyocytes.