Promoter, recombined mesenchymal stem cells and preparing method and application of recombined mesenchymal stem cells
A technology of mesenchymal stem cells and promoters, applied in the field of biomedicine, can solve the problems of poor curative effect of osteoarthritis and achieve good therapeutic effect
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Embodiment 1
[0039] The present invention provides a promoter, the sequence of which is shown in SEQ ID NO:2. The promoter can positively activate and express the TNFα antibody gene in the presence of the inflammatory factor TNFα, and the recombinant mesenchymal stem cells for treating osteoarthritis can be obtained by using the promoter.
[0040] The promoter provided by the present invention positively activates and expresses the TNFα antibody gene in the presence of the inflammatory factor TNFα. The mechanism of action is: the inflammatory factor TNFα has multiple downstream signaling pathways, and the most important downstream signaling pathway is NFα Activation of -κB transcription factor; while the inflammatory factor TNFα activates the NF-κB pathway, many signaling molecules are involved, such as TNFR-related factors, receptor-interacting protein (RIP, Receptor-interacting protein), mitogen-activated protein kinase 3 ( MAP3K, mitogen-activated protein kinase ki-nase 3), IKK complex,...
Embodiment 2
[0042] Such as figure 1 Shown, a preparation method of recombinant mesenchymal stem cells, comprising:
[0043] Step S1: providing the TNFα antibody gene and the above-mentioned promoter, and constructing a lentiviral expression plasmid; wherein, the sequence of the TNFα antibody gene is shown in SEQ ID NO:1;
[0044] Step S2: packaging the lentiviral expression plasmid with lentivirus to obtain virus liquid;
[0045] Step S3: Infect the mesenchymal stem cells in the virus fluid to obtain the desired recombinant mesenchymal stem cells. Among them, mesenchymal stem cells (MSC) can be purchased directly, or extracted from animal bone marrow. In the specific operation of the present invention, MSCs are extracted from the bone marrow of C57 / BL6 mice, and the C57 / BL6 mice are purchased from the Experimental Animal Center of the University of Hong Kong.
[0046] The recombinant mesenchymal stem cells are prepared by using the above-mentioned promoters. The recombinant mesenchymal...
Embodiment 3
[0078] A recombinant mesenchymal stem cell integrated with a promoter and a TNFα antibody gene; the sequence of the promoter is shown in SEQ ID NO:2, and the sequence of the TNFα antibody gene is shown in SEQ ID NO:1. That is, it is prepared by adopting the above-mentioned preparation method of recombinant mesenchymal stem cells. The recombinant mesenchymal stem cells are integrated with the promoter and TNFα antibody gene, which will be activated by the inflammatory factor TNFα in an inflammatory environment, and can stably bind to the inflammatory site, thereby enhancing the anti-inflammatory and immune regulation capabilities of the mesenchymal stem cells , so it can be used to treat osteoarthritis with good therapeutic effect.
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