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[Construction of Rat interleukin-10 Adenoviral Vector and Its Expression in Bone Marrow Mesenchymal Stem Cells]

[Construction of Rat interleukin-10 Adenoviral Vector and Its Expression in Bone Marrow Mesenchymal Stem Cells]

Wen-Ting Zhang, Juan Tang, Hong-Mei Zhao, Jie-Yu You

Zhongguo Dang Dai Er Ke Za Zhi. 2019 Jul;21(7):708-712.

PMID: 31315773

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42413339	Interleukin-10 from rat			Price

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