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Spleen mesenchymal stem cell with immune adjustment function, preparation method of cell and application of cell

A mesenchymal stem cell and spleen technology, applied in the field of spleen mesenchymal stem cells and their preparation, can solve problems affecting scientific research and application, removal, etc., and achieve the effect of inhibiting T lymphocyte proliferation, high purity, and simple operation

Pending Publication Date: 2017-01-04
CHINESE PEOPLES LIBERATION ARMY AIR FORCE GENERAL HOSPITAL
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  • Application Information

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Problems solved by technology

However, when this strategy is applied to the culture of Sp-MSCs, it encounters difficulties. The reason is that the spleen is rich in a large number of hematopoietic cells such as macrophages, which have a strong ability to attach to culture bottles and are difficult to pass passage. culture removal
Hematopoietic cells doped in Sp-MSCs often cause immune responses, seriously affecting related scientific research and applications

Method used

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  • Spleen mesenchymal stem cell with immune adjustment function, preparation method of cell and application of cell
  • Spleen mesenchymal stem cell with immune adjustment function, preparation method of cell and application of cell
  • Spleen mesenchymal stem cell with immune adjustment function, preparation method of cell and application of cell

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Experimental program
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Embodiment 1

[0067] Embodiment 1, a kind of method for separating and cultivating Sp-MSCs

[0068] 1. Isolation and culture of Sp-MSCs

[0069] 1. Separation of the spleen

[0070] Healthy female C57 mice aged 6-8 weeks were killed by neck dislocation. After soaking in 75% alcohol for 3-5 minutes, the spleen was isolated under aseptic conditions. The surface tissue was removed with scissors and tweezers, and the complete spleen was removed with tweezers.

[0071] 2. Removal of hematopoietic cells in the spleen

[0072] Fully grind it in a culture dish with a syringe handle for 5 minutes in an ultra-clean bench, rinse it with PBS 3-5 times, remove most of the hematopoietic cells in the spleen, and obtain the spleen stromal tissue ( figure 1 A).

[0073] 3. Collagenase digestion of spleen matrix tissue and primary culture

[0074] Place the spleen matrix tissue isolated in step 2 into collagenase solution for digestion (put 3-5 mouse spleen matrix tissues in one milliliter of collagenase...

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Abstract

The invention discloses a spleen mesenchymal stem cell with an immune adjustment function, a preparation method of the cell and an application of the cell. By the aid of the strategy of removing hematopoietic cell-digestion matrix tissues to obtain Sp-MSCs-passage purification, MSCs are cultured in an isolated manner from the spleen, and a large number of Sp-MSCs with high cell purify are obtained by isolation. Further, the Sp-MSCs have high immune adjustment function, heterogenetic antigen and homologous antigen activated T lymphocyte proliferation and activation can be inhibited, and inflammatory cytokine secretion of T lymphocyte is inhibited.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a spleen mesenchymal stem cell with immune regulation function and its preparation method and application. Background technique [0002] Mesenchymal stem cells (MSCs) are stem cells with multilineage differentiation potential and high self-renewal ability. MSCs were first discovered in bone marrow by Friedenstein et al., and then researchers successively isolated them from bone, umbilical cord, fat and other tissues. Under suitable conditions, MSCs can be expanded in large quantities and differentiated into cells of various tissue types such as osteoblasts, adipocytes, endothelial cells, and nerve cells. They are currently one of the most commonly used seed cells in the field of tissue engineering. In addition, MSC also has the ability to support hematopoiesis, can regulate the differentiation of hematopoietic stem cells into hematopoietic cells of various lineages, and is an import...

Claims

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Application Information

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IPC IPC(8): C12N5/0775C12N5/0783
CPCC12N5/0668C12N5/0636C12N2509/00
Inventor 王恒湘丁丽郭子宽朱恒
Owner CHINESE PEOPLES LIBERATION ARMY AIR FORCE GENERAL HOSPITAL
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