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Separation method of tree shrew peripheral blood mononuclear cells

A separation method and peripheral blood technology, applied in the field of cell separation, can solve problems such as hindering the study of immune cells of tree shrews, harming animal welfare, and increasing the demand for blood collection of tree shrews

Pending Publication Date: 2021-11-05
THE FIRST AFFILIATED HOSPITAL OF GUANGXI MEDICAL UNIV
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Problems solved by technology

[0004] However, in the application of methods and reagents in the prior art, there are no reference methods and corresponding reagents for the separation of tree shrew PBMCs. In actual work, researchers have to select the separation liquid of other species for separation, but due to the differences between species The concentration and purity of tree shrew PBMCs obtained are not ideal, which will increase the demand for blood collection of tree shrews and damage animal welfare; on the one hand, it will also hinder further research on tree shrew immune cells

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  • Separation method of tree shrew peripheral blood mononuclear cells
  • Separation method of tree shrew peripheral blood mononuclear cells
  • Separation method of tree shrew peripheral blood mononuclear cells

Examples

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Embodiment 1

[0032] Example 1: Establishing the method and verification of isolating mononuclear cells from the peripheral blood of experimental tree shrews

[0033] 1 Materials and methods

[0034] 1.1 The experimental animals were adult tree shrews of the Burmese species artificially bred and cultured in the Experimental Animal Center of Kunming Medical University. They were raised in the common tree shrew breeding room of the Experimental Animal Center of Guangxi Medical University. They were reared in single cages and the indoor temperature was (24 ± 1) ℃, relative humidity 40% to 50%, artificial lighting 12h / d. Regularly clean the feces every day, feed nutritious mealworms and fruits, and drink water freely. The animal experiments in this study complied with the "3R" principle.

[0035] 1.2 Main reagents and instruments Separation solution (Sigma, USA); Hanks balanced salt solution (Solarbio, Beijing); Whole blood and tissue diluent (Solarbio, Beijing); Red blood cell lysate (×10)...

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Abstract

The invention discloses a separation method of tree shrew peripheral blood mononuclear cells, and mainly relates to the technical field of cell separation. The separation method comprises the following steps of: separating diluted anticoagulant whole blood cell components of tree shrew by adopting a gradient centrifugation method of separation solutions with different densities, and obtaining PBMC. The separation method has the beneficial effects that 1, the proper separation density of the PBMC of the tree shrew and a corresponding separation solution preparation method are provided, so that the PBMC of the tree shrew with large quantity and high purity can be quickly obtained; and 2, when all white blood cell (including the PBMC) components in the peripheral blood of the tree shrew need to be obtained, the effect evaluation of direct lysis of the peripheral blood of the tree shrew by using red blood cell lysis solutions with different volumes is also given, and reference is provided for acquisition and related research of immune cells of the peripheral blood of the tree shrew.

Description

technical field [0001] The invention relates to the technical field of cell separation, in particular to a method for separating peripheral blood mononuclear cells of tree shrews. Background technique [0002] The tree shrew (Tupaia Belangeris) is a small mammal similar to a squirrel, belonging to the small primates, mainly found in South and Southeast Asia. According to the genetic analysis of the Kunming Institute of Zoology, Chinese Academy of Sciences, it was found that tree shrews are highly similar to humans in terms of nervous system and immune system. Studies have shown that the immune response of tree shrews is closer to that of humans than that of rodents. When exposed to a variety of human pathogens, tree shrews exhibit clinical symptoms similar to human infection. [0003] Peripheral blood mononuclear cells (PBMCs), including lymphocytes and monocytes, are the most common cell model in cellular immunology research. Using tree shrews as an experimental animal m...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078
CPCC12N5/0634C12N2509/10C12N2509/00
Inventor 唐安洲夏巍陈泓霖李骅冯清源
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGXI MEDICAL UNIV
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