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Mesenchymal stem cell treating method for Schistosoma japonicum infection treatment

A technology of schistosomiasis and cells, applied in animal cells, anti-infective drugs, vertebrate cells, etc., can solve the problems that there is no MSC to treat Schistosoma japonicum infection, restrict the treatment of Schistosoma japonicum infection, etc., achieve good application prospects and promotion value, alleviate Hepatic fibrosis, good curative effect

Active Publication Date: 2020-02-11
SUN YAT SEN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Therefore, there is no specific plan for using MSCs to treat Schistosoma japonicum infection, which restricts the treatment of Schistosoma japonicum infection.

Method used

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  • Mesenchymal stem cell treating method for Schistosoma japonicum infection treatment
  • Mesenchymal stem cell treating method for Schistosoma japonicum infection treatment
  • Mesenchymal stem cell treating method for Schistosoma japonicum infection treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The isolation culture of embodiment 1 MSC

[0034] 1. Experimental method

[0035] Take about 5-6W mice, kill them by cervical dislocation, soak them in 75% ethanol for 5 minutes, take out both femurs under aseptic condition, carefully remove the attached tissues on the femurs, and use a syringe to draw 100mL / L-DMEM medium of L FCS, wash out the cells in the bone marrow cavity, collect the cells through a 200-mesh stainless steel mesh, dissolve the red blood cells in the erythrocyte lysate for 2 minutes, centrifuge to remove the supernatant, and use L-DMEM medium containing 100m L / L FCS Resuspend and adjust cell concentration, 3ⅹ10 4 Inoculated in 24-well plates, placed at 37°C, with a volume fraction of 5% CO 2 In the incubator, change the medium for the first time after 7 days, and then change the medium once every 3 days. When the cells have cell clones, digest and collect the cell clones, transfer them to the culture flask, change the medium every 3 days, and wai...

Embodiment 2

[0038] The processing of embodiment 2 MSC

[0039] 1. Experimental method

[0040]Add IFN-γ (5 ng / ml) and LPS (100 ng / ml) to the MSCs cultured in Example 1, act at 37° C. for 24 hours, 1500 rpm, centrifuge for 10 minutes, discard the culture medium, resuspend with PBS, and centrifuge to remove the supernatant. Repeat the wash once and collect the cells for later use.

[0041] The induced MSC cells were collected, mRNA was extracted, real-time PCR was used to quantitatively detect the expression of the key immune regulatory factor iNOS, and the lymphocyte proliferation inhibition experiment was further performed.

[0042] 2. Experimental results

[0043] After synergistic induction with lower concentration of IFN-γ and lower concentration of LPS, it can promote the expression of MSC iNOS and enhance the inhibitory effect on lymphocyte proliferation, showing that its immune regulation effect is enhanced, such as figure 2 shown.

Embodiment 3

[0044] The induced MSCs of Example 3 are used for the treatment of hepatic fibrosis infected by schistosomiasis

[0045] 1. Treatment

[0046] 1. Take the cercariae of schistosomiasis and infect the mice percutaneously to establish the schistosomiasis infection model;

[0047] 2. MSC treatment

[0048] The schistosome infection model was randomly divided into three groups (each group) with 16 ± 2 cercariae of schistosomiasis, without any treatment as the blank control group, and the other two groups were given tail vein injection of MSC prepared in Example 2 respectively at 2 and 4 weeks after infection. ; Uninduced MSCs were used as the control group, 1×10 5 / per mouse;

[0049] 3. Samples were collected and analyzed 8 weeks after infection.

[0050] 1) The livers of the three groups of schistosome cercariae were collected for HE and masson staining and the quantification of hydroxyproline to analyze the effect of MSC on liver granuloma and fibrosis.

[0051] 2) The seru...

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Abstract

The invention discloses a mesenchymal stem cell treating method for Schistosoma japonicum infection treatment. IFN-gamma and LPS are added into an MSC culture medium to perform cultivation for 24 h at37 DEG C, the final concentration of the IFN-gamma is 5-100 ng / ml, and the final concentration of the LPS is 10-100 ng / ml; and stimulating factors are removed, and therefore, treatment can be realized after full washing. The advantages of the method are mainly manifested as follows: 1, compared with treatment mainly aiming at parasites at present, optimized and induced MSCs can be used for the treatment of the hepatic fibrosis of infected patients and have no obvious toxic and side effects; and 2, compared with the MSCs that are not induced, the optimized and induced MSCs can significantly reduce the hepatic fibrosis caused by schistosome infection by obviously promoting Th1 reaction, so that better curative effect can be obtained.

Description

technical field [0001] The present invention relates to the technical field of parasitic infection treatment, more specifically, relates to a treatment method of mesenchymal stem cells for the treatment of Schistosoma japonicum infection. Background technique [0002] Schistosomiasis is a zoonotic parasitic disease caused by schistosome infection that seriously affects human health and social and economic development. The schistosomiasis parasitic on the human body mainly include Schistosoma japonicum, S. S. haematobium), S. intercalatum, S. mekongi and S. malayensis. According to the statistics of the World Health Organization (WHO), about 200 million people in the endemic areas of 74 countries are still affected and threatened by schistosomiasis. According to statistics, as of the end of 2016, there were still about 54,000 schistosomiasis patients in my country, and the prevention and control situation in some areas was still grim. In my country, the main pathogen that c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61K35/28A61P33/12
CPCA61K35/28A61P33/12C12N5/0663C12N2501/052C12N2501/24Y02A50/30
Inventor 雷俊霞吴忠道
Owner SUN YAT SEN UNIV
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