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Method for generating myeloid suppressor cells through sodium urate induction[v]

A technology of suppressive cells and sodium urate, which is applied in the field of biomedicine, can solve the problems of small number of cells and in vitro expansion methods that have not been described in detail, and achieve the effect of good application prospects

Active Publication Date: 2020-05-26
GUANGDONG NO 2 PROVINCIAL PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although major breakthroughs have been made in the field of myeloid-derived suppressor cells, due to the small number of myeloid-derived suppressor cells, the safe, non-tumor-derived myeloid-derived Methods for in vitro expansion of suppressor cells are not yet fully described

Method used

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  • Method for generating myeloid suppressor cells through sodium urate induction[v]
  • Method for generating myeloid suppressor cells through sodium urate induction[v]
  • Method for generating myeloid suppressor cells through sodium urate induction[v]

Examples

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Effect test

Embodiment 1

[0034] Example 1 sodium urate transforms mouse monocytes into myeloid suppressor cells (MDSCs)

[0035] 1. Experimental method

[0036] In this embodiment, a method for inducing myeloid suppressor cells by sodium urate comprises the following steps:

[0037] (1) Isolation of monocytes from mouse bone marrow:

[0038] 1) Kill 6-week-old C57 mice by cervical dislocation, immerse the mice in 75% alcohol for 3 minutes to disinfect; take out the mice, cut the skin on both sides of the mice with ophthalmic scissors, and cut off both sides of the legs from the hip joint bone. Carefully cut off the attached muscles of the femur and tibia, without injuring the articular surface, and avoid exposing the bone marrow cavity;

[0039] 2) In a biological safety cabinet, soak the stripped femur and tibia in 75% alcohol for 3 minutes and transfer them to pre-cooled RPMI 1640 medium;

[0040] 3) Gently cut open both ends of the femur or tibia with ophthalmic scissors, then use a 5ml sterile...

Embodiment 2

[0069] Example 2 Sodium urate converts human peripheral blood mononuclear cells into myeloid suppressor cells (MDSCs)

[0070] 1. Experimental method

[0071] In this embodiment, a method for inducing myeloid suppressor cells by sodium urate comprises the following steps:

[0072] (1) Isolation of mononuclear cells from human peripheral blood:

[0073] Take 5ml of peripheral blood from healthy volunteers and separate mononuclear cells;

[0074] The specific method is as follows:

[0075] 1) Use EDTA-containing anticoagulant blood collection tubes to collect peripheral blood from healthy volunteers, then add 3 times the volume of PBS solution, and mix gently;

[0076] 2) Add 3ml of lymphocyte separation solution to a 50ml sterile centrifuge tube; then use a pipette to slowly add the blood dilution solution to the upper layer of the lymphocyte separation solution along the tube wall, at 18°C, 600g (liter 5, drop 0) , centrifuge for 22min;

[0077] 3) After centrifugation, s...

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Abstract

The invention discloses a method for generating myeloid suppressor cells through sodium urate induction. The method comprises the following steps: (1) separating mononuclear cells from a sample; (2) carrying out in-vitro culture on the monocytes obtained in the step (1) by using a complete culture medium containing GM-CSF; and (3) adding sodium urate into the culture system in the step (2), and continuously carrying out in-vitro culture to obtain the myeloid suppressor cells. The method is safe and simple, the myeloid suppressor cells with the immunosuppressive function can be generated through rapid induction, and the myeloid suppressor cells prepared through the method can be used for treating autoimmune diseases, neoplastic diseases and the like and have good application prospects.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for inducing myeloid suppressor cells produced by sodium urate. Background technique [0002] Myeloid-derived suppressor cells (MDSCs) are a heterogeneous cell population with immunosuppressive function initially identified in tumor-bearing mice. Tumor cells regulate the tissue microenvironment in bone marrow and spleen by releasing soluble factors that can promote the accumulation of myeloid cells. Normal differentiation pathways develop into terminal differentiation of mature macrophages, dendritic cells, and granulocytes shifting to the accumulation of immature myeloid cells differentiated in a highly immunosuppressed state, which have been named myeloid suppressor cells to highlight their Common myeloid origin and immunomodulatory properties. Immature myeloid cells (IMCs) with the same phenotype as MDSCs are continuously generated and differentiated...

Claims

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Application Information

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IPC IPC(8): C12N5/078
CPCC12N5/0634C12N2500/30
Inventor 曹东林钟丽梅
Owner GUANGDONG NO 2 PROVINCIAL PEOPLES HOSPITAL
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