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Transgenic mesenchyma stem cell for curing radiation enteritis and method of producing the same

A radiation enteritis, stem cell technology, applied in botanical equipment and methods, biochemical equipment and methods, cells modified by introducing foreign genetic material, etc. Easy to obtain, low immunogenicity, and anti-apoptotic effect

Inactive Publication Date: 2008-05-21
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Studies by Devine et al. have shown (Devine SM, et al. Mesenchymal stem cells distribute to a wide range of tissues following systemic infusion into nonhuman primates. Blood, 2003, 101(8): 2999-3001.), vein transplanted MSCs can homing Colonizes the intestinal tract, but the number is very small, accounting for only about 1-2% of the recipient intestinal mucosal cells, unable to play a role in repair and regeneration

Method used

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  • Transgenic mesenchyma stem cell for curing radiation enteritis and method of producing the same
  • Transgenic mesenchyma stem cell for curing radiation enteritis and method of producing the same
  • Transgenic mesenchyma stem cell for curing radiation enteritis and method of producing the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Preparation method of transgenic mesenchymal stem cells

[0026] (1) Isolation and culture of MSCs:

[0027] Isolation and culture of bone marrow MSCs: Take the iliac bone marrow puncture of the donor, add heparin for anticoagulation, mix with DMEM culture medium at a ratio of 1:5, centrifuge at 200×g for 10 minutes, remove the bone residue and place in 1.077×10 3 centrifuge at 400×g for 20 min to collect mononuclear cells, wash twice with 0.1mol / L phosphate buffer, and wash with 4×10 5 / cm 2 Inoculate in a culture flask at a density of 37°C, 5% CO 2 Cultivate in an incubator, remove non-adherent cells after 72 hours, change the medium twice a week, and digest with 0.25% trypsin when the cells grow to 90% confluence at the bottom of the bottle, passage 1:3, and take the third generation cells for the following steps;

[0028] Isolation and culture of adipose tissue MSCs: The adipose tissue was thoroughly washed three times with 0.1mol / L phosphate buffer t...

Embodiment 2

[0035] Example 2: Method of using transgenic mesenchymal stem cells

[0036] The transfected mesenchymal stem cells were digested with 0.25% trypsin, blown into a single-cell suspension with 0.1mol / L phosphate buffer, and the concentration was adjusted to 5×10 7 / ml, transplanted by intravenous injection, the dose is 1×10 9 cells / time, once every other day, a total of 3 injections.

Embodiment 3

[0037] Embodiment 3: animal experiment research

[0038] The following is an animal experiment study on the treatment of radiation enteritis by transgenic mesenchymal stem cells to prove the experimental basis and actual effect of the present invention.

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Abstract

The invention discloses a transgenic mesenchymal stem cell which can treat radiation enteritis and a preparation method. The CXCR4 protein can be expressed highly on the surface of transgenic mesenchymal stem cell; under the chemotaxis of stromal cell derived factor which is released from the injured intestine, the homing and colonization of a large amount of transgenic mesenchymal stem cells can promote proliferation and repair of the intestinal mucosa and inhibit apoptosis; the radiation enteritis is cured. The steps of the preparation method of the transgenic mesenchymal stem cell are that : (1)the separation and cultivation of mesenchymal stem cell; (2) the preparation of CXCR4 recombinant adenovirus: full length cDNA of CXCR4 is amplified by RT-PCR; the construction of CXCR4 recombinant shuttle plasmid and skeleton plasmid; the CXCR4 recombinant adenovirus is packaged, amplified, purified and determined by titer in HEK293T cell; (3) the transfect ion of mesenchymal stem cell by using CXCR4 recombinant adenovirus.

Description

1. Technical field [0001] The invention relates to the field of biological preparations, in particular to a transgenic mesenchymal stem cell capable of treating radiation enteritis and a preparation method thereof. The CXCR4 protein is highly expressed on the surface of the cells, which can homing and colonize a large number of radiation-damaged intestinal mucosa, promote proliferation and repair, and inhibit apoptosis, thereby treating radiation enteritis. 2. Background technology: [0002] Radiation enteritis is the most common complication during clinical radiotherapy for abdominal tumors. According to the statistics of the World Health Organization (WHO), there are more than 15 million people suffering from malignant tumors every year in the world, and more than 60% of the patients need to use radiotherapy as the main or auxiliary treatment during the treatment process. Radiation intestinal injury occurs in about 5-10% of patients after abdominal radiotherapy, mainly ma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/861A61K35/12A61P35/00A61P1/00
Inventor 张坚朱维铭王祖森李秋荣黎介寿
Owner NANJING UNIV
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