关键词: 骨髓间充质干细胞')">骨髓间充质干细胞, 5-氮杂胞苷, 心肌细胞

Abstract:

【Objective】 This research aimed to explore 5-Aza inducing BMSC into cadiocyte utilizing molecular biology technology so as to provide seed cells for genetically modified varieties of beef cattle and elevating transgenic efficiency. 【Method】 Test is divided into three groups，they were the P4, P8, P12 BMSC of purification and stability. Each group utilized L-DMEM containing 5, 10, 15 μmol?L-1 5-Aza to undertake respectively derivation，statistics and calculate transformation efficiency of cadiocyte, accredit cadiocyte by observation of morphology and RT-PCR.【Result】 Most cells after differentiation showed fusiform shape, orientating with one accord, forming myotubule-like structure. The inductivity of same passage BMSC was the highest in 10 μmol?L-1 5-Aza role.The conversion rate of BMSC to cadiocyte decreased with the increasing passage number at the same concentration of 5-Aza. RT-PCR results showed that cells after induction clearly expressed cardiac four specific genes, they are α-actin (378 bp), desmin (601 bp), cardiac troponin T (331 bp), β-myosin heavy chain (β -MHC) (273 bp). 【Conclusion】 5-Aza can induce bovine bone marrow mesenchymal stem cells into cadiocyte in vitro. Effect of derivation is obvious utilizing P4 BMSC with 10 μmol?L-1 5-Aza.

Key words: bone marrow mesenchymal stem cells')">bone marrow mesenchymal stem cells, 5-azacytidine, cadiocyte