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CJCP  2019, Vol. 21 Issue (7): 708-712    DOI: 10.7499/j.issn.1008-8830.2019.07.017
EXPERIMENTAL RESEARCH Current Issue| Next Issue| Archive| Adv Search  | 
Construction of rat interleukin-10 adenoviral vector and its expression in bone marrow mesenchymal stem cells
ZHANG Wen-Ting1, TANG Juan2, ZHAO Hong-Mei1, YOU Jie-Yu1
Department of Gastroenterology, Hunan Children's Hospital, Changsha 410007, China
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Abstract  Objective To construct the recombinant adenoviral vector carrying the rat interleukin-10 (rIL-10) gene, and to investigate whether it is stably expressed in bone marrow mesenchymal stem cells. Methods The rIL-10 gene was amplified by PCR from template rIL-10 cDNA, and the recovered 656 bp rIL-10 DNA fragment was cloned into pcDNA3.1 to construct pcDNA3.1-IL-10. Then HEK293 cells were transfected with pcDNA3.1-IL-10 and adenoviral vector for homologous recombination, and sequencing and PCR were used to evaluate whether recombination was successful. HEK293 cells were lysed by repeated freeze-thaw cycles, and bone marrow mesenchymal stem cells were infected with the virus solution containing the rIL-10 gene. Western blot was used to measure the expression of rIL-10 in bone marrow mesenchymal stem cells. Results Sequencing and PCR verified that the rIL-10 adenoviral vector was successfully constructed, with a virus titer of 4×109 PFU/mL. The expression of IL-10 was detected after bone marrow mesenchymal stem cells were infected by the virus solution containing the rIL-10 gene. Conclusions The constructed rIL-10 recombinant adenovirus can mediate the stable expression of rIL-10 gene in bone marrow mesenchymal stem cells, which provides a basis for gene transplantation therapy of inflammatory bowel disease.
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ZHANG Wen-Ting
TANG Juan
ZHAO Hong-Mei
YOU Jie-Yu
Key wordsInflammatory bowel disease      Interleukin-10 gene      Adenoviral vector      Bone marrow mesenchymal stem cell      Rats     
Received: 25 December 2018     
Cite this article:   
ZHANG Wen-Ting,TANG Juan,ZHAO Hong-Mei et al. Construction of rat interleukin-10 adenoviral vector and its expression in bone marrow mesenchymal stem cells[J]. CJCP, 2019, 21(7): 708-712.
ZHANG Wen-Ting,TANG Juan,ZHAO Hong-Mei et al. Construction of rat interleukin-10 adenoviral vector and its expression in bone marrow mesenchymal stem cells[J]. CJCP, 2019, 21(7): 708-712.
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