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Gene modified mesenchymal stem cell and application thereof

A technology of mesenchymal stem cells and genetic modification, applied in the field of genetically modified cells and tumor treatment, can solve the problems of limiting the role of MSCs, lack of sufficient evidence to show MSCs, and accelerate the development of tumors, etc., to achieve the effect of prolonging the survival period

Pending Publication Date: 2020-12-04
深圳市芥至和生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in a specific tumor microenvironment, because MSCs have a strong immune regulation effect, MSCs may promote the proliferation of tumors, thereby accelerating the occurrence and development of tumors, which greatly limits the role of MSCs in tumor therapy.
[0011] In conclusion, there is no sufficient evidence to show whether MSCs promote or inhibit tumor proliferation, and there is no report on the combination of MSCs and chemokines to treat or prevent tumors.

Method used

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  • Gene modified mesenchymal stem cell and application thereof
  • Gene modified mesenchymal stem cell and application thereof
  • Gene modified mesenchymal stem cell and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Preparation of genetically modified mesenchymal stem cells

[0058] 1. Construction of lentiviral plasmid vector

[0059] 1) Synthesize and amplify the nucleotide sequences SEQ ID NO:3 and SEQ ID NO:4 encoding the CCL5 and CXCL9 proteins of the present invention, completed by Shanghai Langjing Biotechnology Co., Ltd.;

[0060] 2) Cloning SEQ ID NO: 3 and SEQ ID NO: 4 into the lentiviral expression vector pGreen puro respectively by conventional genetic engineering means to obtain the lentiviral expression vector A containing the nucleotide sequence encoding the CCL5 protein and the lentiviral expression vector A containing the encoding CXCL9 The lentiviral expression vector B of the nucleotide sequence of the protein.

[0061] The specific process of constructing lentiviral expression vector A and lentiviral expression vector B is as follows:

[0062] Insert SEQ ID NO: 3 and SEQ ID NO: 4 into the lentiviral expression vector pGreen puro separately through th...

Embodiment 2

[0089] Example 2 Verification of the effect of the gene-modified mesenchymal stem cells provided by the present invention in treating tumors (breast cancer, pancreatic cancer, liver cancer, lung cancer, gastric cancer, colon cancer) in vivo

[0090] (1) Take 15 female nude mice (6 weeks old, weighing 18-20g, purchased from Guangdong Medical Experimental Animal Center), and inject 5x 10 6 MCF-7 (breast cancer) cells, when the tumor grows to 60mm 3 Size, tumor models were randomly divided into 3 groups:

[0091] a. Control group: intravenous injection of normal saline 200ul / time, once a week, 4 times in total;

[0092] b. hUC-MSCs cell therapy group: 3×10 hUC-MSCs cells were injected into the tail vein 6 One piece / time, 200ul in total, once a week, 4 times in total;

[0093] c.hUC-MSCs-CCL5-CXCL9 cell therapy group: tail vein injection

[0094] hUC-MSCs-CCL5-CXCL9 cells 3×10 6 Each time, a total of 200ul, once a week, a total of 4 times; the survival status of the mice with...

Embodiment 3

[0128] Example 3 Verification of the in vivo tumor prevention effect of the genetically modified mesenchymal stem cells provided by the present invention (1) Take 15 female nude mice (6 weeks old, weighing 18-20 g, purchased from Guangdong Medical Experimental Animal Center), and randomly divide them into 3 groups:

[0129] a. Control group: Inject normal saline 200ul / time into tail vein, once a week, 4 times in total;

[0130] b. hUC-MSCs cell prevention group: 3×10 hUC-MSCs cells were injected into the tail vein 6 One piece / time, 200ul in total, once a week, 4 times in total;

[0131] c.hUC-MSCs-CCL5-CXCL9 cell prevention group: tail vein injection

[0132] hUC-MSCs-CCL5-CXCL9 cells 3×10 6 Each time, a total of 200ul, once a week, a total of 4 times.

[0133] Afterwards, the mice in each group were subcutaneously injected with 5x 10 6 For MCF-7 (breast cancer) cells, the survival status of the mice within 120 days was counted, and the survival rate curve was made. The ...

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Abstract

The invention provides a gene modified mesenchymal stem cell and application thereof. The gene modified mesenchymal stem cell expresses CCL5 and CXCL9 proteins. The invention also provides applicationof the gene modified mesenchymal stem cell in preparation of drugs for treating or preventing tumors, and application of the gene modified mesenchymal stem cell in preparation of vaccines for treating or preventing tumors. The scheme for treating or preventing tumors by using the gene modified mesenchymal stem cell provided by the invention can significantly prolong the lifetime of tumor-bearingmice.

Description

technical field [0001] The invention relates to the technical field of genetically modified cells and tumor treatment, in particular to genetically modified mesenchymal stem cells and applications thereof. Background technique [0002] Tumors are a serious threat to human health. In China, the incidence of tumors is increasing year by year, and the mortality rate of tumors is increasing rapidly. Malignant tumors rank second among various causes of death nationwide, and rank first in urban areas. . How to reduce the incidence of tumors and prolong the survival of cancer patients has become the focus of researchers in recent years. Compared with surgical intervention and chemotherapy, tumor immunotherapy has significantly reduced side effects and strong targeting. current focus. [0003] Tumor immunotherapy is a treatment method to control and eliminate tumors by restarting and maintaining the tumor-immune cycle and restoring the body's normal anti-tumor immune response. Th...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/867C12N15/12A61K35/28A61K39/00A61P35/00
CPCC12N5/0662C12N15/86C07K14/522C07K14/523A61K35/28A61K39/001142A61P35/00C12N2740/15043C12N2800/107
Inventor 刘未斌刘曲波李琼书
Owner 深圳市芥至和生物科技有限公司
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