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Serum-free culture fluid for enhancing immunosuppression function of animal mesenchymal stem cells

A serum-free culture medium and serum-free culture technology, applied in animal cells, vertebrate cells, bone/connective tissue cells, etc., can solve the limited nutritional promotion effect of animal mesenchymal stem cells, without considering the impact of immunosuppressive effects , poor reproducibility of products and experimental results, etc., to achieve the effect of favoring differentiation, stabilizing signal transduction pathways, and avoiding toxic effects

Inactive Publication Date: 2017-01-04
周宇璠 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Use animal mesenchymal stem cell serum culture medium, which is expensive;
[0008] Due to different production batch numbers, there will be differences between batches, resulting in poor reproducibility of products and experimental results;
[0009] The protein components brought in by the serum itself need to be removed, and if necessary, purification is required, which complicates the operation;
[0010] Unknown pathogens (viruses, etc.) may be mixed in the serum, which may contaminate the cultured cells
[0011] The publication number is CN102703385A, and the publication date is October 3, 2012. The Chinese invention patent application titled "a culture medium for mesenchymal stem cells" discloses a serum-free culture medium for animal mesenchymal stem cells; and the publication number CN103805562A, published on May 21, 2014, the Chinese invention patent application titled "Serum-free medium for cultivating placental mesenchymal stem cells" discloses a serum-free culture for culture and expansion of placental mesenchymal stem cells However, the composition of the serum-free medium of these inventions is relatively simple, and the nutritional promotion effect on the growth and expansion of animal mesenchymal stem cells is limited
[0012] None of the aforementioned patents involving mesenchymal stem cell culture fluid took into account the impact on the immunosuppressive effect of the cultured mesenchymal stem cells, which may reduce the immunosuppressive effect of the cultured mesenchymal stem cells

Method used

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  • Serum-free culture fluid for enhancing immunosuppression function of animal mesenchymal stem cells
  • Serum-free culture fluid for enhancing immunosuppression function of animal mesenchymal stem cells
  • Serum-free culture fluid for enhancing immunosuppression function of animal mesenchymal stem cells

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Effect test

Embodiment 1

[0052] Human bone marrow-derived mesenchymal stem cells were co-cultured with mouse-derived activated T cells, and the culture medium used was respectively 10% FBS-MSCGM medium and animal mesenchymal stem cell serum-free medium (SF-MSCGM) of the present invention for cultivation. The composition of the invented SF-MSCGM is as follows: basal culture medium + proliferation factors (bFGF, HGF, TGF-β3, PDGF-BB) + CD + cholesterol + VE + lecithin + dexamethasone + transferrin + selenous acid + BSA + insulin + INF -γ+TNF-α. For the T cell proliferation response stimulated by anti-CD3 and CD28, the results show that the human mesenchymal stem cells cultivated with the serum-free medium of the present invention have an immunosuppressive effect ( Figure 1-a , Figure 1-b ).

Embodiment 2

[0054] Take human bone marrow-derived mesenchymal stem cells and mouse-derived activated T cells for co-cultivation, and the culture medium used is respectively 10% FBS-MSCGM culture medium and animal mesenchymal stem cell immunosuppression of the present invention can enhance serum-free culture medium (SF-MSCGM) , The composition of SF-MSCGM of the present invention is the same as in Example 1. For the T cell proliferation response stimulated by anti-PMA / Ionomyein, the results show that human mesenchymal stem cells cultivated with serum-free medium of the present invention have immunosuppressive effect ( Figure 2-a , Figure 2-b ).

Embodiment 3

[0056] Experiment of immunosuppressive effect of hMSC derived from bone marrow on T cell proliferation under mixed lymphocyte reaction (MLR) of mice. The experimental method, the cells used, and the culture medium used are all the same as in Example 1. The treatment method of hMSC derived from bone marrow is to sow 2×10 cells in each well of 96 wells. 4 cells at a density; the activation method of mouse spleen cells is to use 2×10 4 Density mouse spleen cells were co-cultured with mouse bone marrow-derived dendritic cells (BMDC), and the seeding density of the latter was 3.3×10 4 pcs / hole. The results of the examples show that the bone marrow-derived hMSCs cultured with SF-MSCGM of the present invention have the same immunosuppressive effect as the bone marrow-derived hMSCs cultured with MSCGM medium; Figure 3-b ), while the bone marrow-derived hMSCs cultured with SF-MSCGM of the present invention began to exhibit immunosuppressive effect from day 3 ( Figure 3-a ). That ...

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Abstract

The invention provides cell culture fluid of serum-free cultivation for enhancing immunosuppression function of animal mesenchymal stem cells. Multiplication factors and specific additives are added into a basic nutrient solution for serum-free cultivation of animal mesenchymal stem cells, in order to promote healthy growth and proliferation of animal mesenchymal stem cells, and provide functions for maintaining and enhancing immunosuppression of animal mesenchymal stem cells. The fluid has the function for enhancing function of animal mesenchymal stem cells. Experiments prove that the serum-free culture fluid for enhancing immunosuppression function of animal mesenchymal stem cells can be used for serum-free cultivation of animal mesenchymal stem cells, and can be applied to clinic research and application in the aspect of immunosuppression application research of mesenchymal stem cells.

Description

technical field [0001] The invention relates to a serum-free culture solution capable of enhancing the immunosuppression of animal mesenchymal stem cells. Proliferation factors and specific additives are added to the basal culture solution for serum-free culture of animal mesenchymal stem cells, which can make animal mesenchymal stem cells The immunosuppressive effect of mesenchymal stem cell proliferation is enhanced. Background technique [0002] In addition to having multiple differentiation potentials, mesenchymal stem cells also have immunoregulatory functions. In vitro experiments have shown that mesenchymal stem cells can inhibit the proliferation of T cells in vitro and maintain T cells in a dormant state instead of apoptosis. Mesenchymal stem cells exhibit a strong inhibitory effect on the immune system after stimulation by inflammatory cytokines. Because mesenchymal stem cells have strong immune regulation function and low immunogenicity, mesenchymal stem cells a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 周宇璠朱轶
Owner 周宇璠
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