Recombinant adenovirus (AdEasy system) mediated exogenous expression of long non-coding RNA H19 (lncRNA H19) biphasic regulating osteogenic differentiation of mesenchymal stem cells (MSCs).
作者： Junyi Liao , Haozhuo Xiao , Guangming Dai , Tongchuan He , Wei Huang
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摘要: Background We previously constructed AdEasy system for rapid generation of recombinant adenovirus expressing coding genes. However, it is unclear if could be used exogenously expression long noncoding RNAs (lncRNAs). Here we investigated how to overexpress lncRNA H19 with and identified the effect overexpression on mesenchymal stem cells (MSCs) osteogenic differentiation. Methods fragment 1 2 were amplified from mouse genomic DNA separately then connected full-length H19. was firstly subcloned homemade pMOK plasmid, cut off pMOK-H19 plasmid. After homologous recombination in AdEasier (BJ5183 cell), packing mammalian packaging cell line amplification 293pTP cells, high titer AdH19 obtained. Immortalized adipose-derived progenitors (iMADs) infected by different infection rate, H19, related microRNAs (miRs) differentiation markers determined TqPCR. Alkaline phosphatase (ALP) activities matrix mineralization ALP assays Alizarin red S staining respectively. Results suitable production can effectively serve as functional (MSCs). Higher dosage inhibited MSCs, however, lower promoted MSCs. Conclusions reported method system, biphasic MSCs
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