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Recombinant mesenchymal stem cells and application thereof

A technology of mesenchymal stem cells and uses, applied in the field of recombinant mesenchymal stem cells, to achieve significant therapeutic effects, low residual, and low risk of immune reactions

Inactive Publication Date: 2019-06-11
广东美赛尔细胞生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Therefore, MSCs play an important role in the occurrence and development of ovarian cancer, which provides a new research idea for the treatment of this disease, but the specific role and mechanism are still controversial, and further research is needed

Method used

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  • Recombinant mesenchymal stem cells and application thereof
  • Recombinant mesenchymal stem cells and application thereof
  • Recombinant mesenchymal stem cells and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1, the preparation method of bone marrow mesenchymal stem cells

[0026] A get 4-week-old male SD rats, and kill them by intraperitoneal injection of pentobarbital sodium;

[0027] B gets the rat femur, cuts off the epiphysis at both ends of the femur with sterile scissors, washes the femur with 5 ml of low-sugar DMEM medium containing 10% fetal bovine serum and 100u / ml of penicillin and streptomycin, and rinses the femur 3 times;

[0028] C repeatedly blowing and blowing the cells with DMEM medium containing 10% fetal bovine serum and 100u / ml of penicillin and streptomycin to make it a single suspension cell;

[0029] D Adjust the cell density to 1.0×10 6 Inoculate in a cell culture flask, replace with a new medium after 3 days, and trypsinize and subculture when the cells reach 80% confluence.

Embodiment 2

[0030] Example 2, Preparation of Recombinant Mesenchymal Stem Cells

[0031] The mRNA gene fragment of IFN-β obtained in 293T cells is integrated into bone marrow mesenchymal stem cells through a lentivirus-mediated method.

Embodiment 3

[0032] Example 3, Proliferation Activity of Recombinant Mesenchymal Stem Cells

[0033] The recombinant mesenchymal stem cells prepared in Example 2 were cultured in low-sugar DMEM medium, and the recombinant mesenchymal stem cells in the logarithmic growth phase were collected respectively, and diluted with low-sugar DMEM medium to 3×10 4 / ml of cell suspension, seeded in 96-well plate. After the recombinant mesenchymal stem cells were fully adhered to the wall, three replicate wells were set up, and a blank control group and a DMSO (dimethyl sulfoxide) solvent control group were set at the same time. After incubating in an incubator for 24h, 48h, 72h, and 96h, add MTT solution and continue to incubate for 1-4h. Use a microplate reader to measure the OD value (optical density value) at 450nm of each well, and measure the absorbance A value. Wherein: the calculation formula of growth inhibition rate is: proliferation rate (%)=(OD test group / OD control group)×100%. According ...

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Abstract

The invention belongs to the technical field of stem cells, and particularly relates to recombinant mesenchymal stem cells and application thereof. According to the recombinant mesenchymal stem cellsprovided by the invention, an mRNA gene segment of IFN-beta obtained from 293T cells is integrated into mesenchymal stem cells; and the prepared recombinant mesenchymal stem cells have the advantagesof being low in immunoreaction risk, high in activity, low in residue and the like, have a remarkable treatment effect on tumor diseases, particularly ovarian cancer, and are ideal preparations for treating the tumor diseases, especially the ovarian cancer diseases.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and in particular relates to a recombinant mesenchymal stem cell and its application. Background technique [0002] More and more theories indicate that the occurrence and development of tumors are related to the tumor microenvironment. Ovarian cancer is the main killer of cancer diseases that endanger women's health. How to treat female ovarian cancer while avoiding the side effects of conventional chemotherapy drugs is a problem that needs to be solved in the treatment of ovarian cancer. In ovarian cancer lesions, chemokines, cytokines, fibroblasts, tumor-associated fibroblasts, mesenchymal stem cells, mononuclear infiltrating cells, T cells, and B cells are commonly found in the tumor microenvironment. Life activities such as proliferation and growth provide raw materials and energy, such as the growth of tumor blood vessels. [0003] Mesenchymal stem cells are a small group of cells deri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10A61K35/28A61P35/00
Inventor 徐单单王一飞
Owner 广东美赛尔细胞生物科技有限公司
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