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Method for preparing single-cell suspension of aortic cells of mouse

A single-cell suspension, aorta technology, applied in cell dissociation methods, animal cells, vertebrate cells, etc., can solve the problems of difficult to obtain single cells in yield, insufficient number of cells, hindering experimental research, etc. The effect of less adhesion and clumping, high single cell yield, and less cell debris

Inactive Publication Date: 2021-01-12
ANHUI NORMAL UNIV
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Problems solved by technology

The preparation of the existing vascular tissue single cell suspension is relatively difficult, and it is difficult to obtain single cells with high yield, high activity and strong function, which hinders further experimental research, and the number of cells is insufficient, cell debris is too much, and cell adhesion Agglomeration, etc. can lead to the failure of the experiment

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  • Method for preparing single-cell suspension of aortic cells of mouse
  • Method for preparing single-cell suspension of aortic cells of mouse
  • Method for preparing single-cell suspension of aortic cells of mouse

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Embodiment Construction

[0024] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some of the embodiments of the present invention, but not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0025] The invention provides a technical solution: a method for preparing a mouse aortic cell single-cell suspension, which specifically includes the following steps:

[0026] S01. First, prepare 1ml of enzyme mixture, surviving healthy wild mice, an operation table with a flat ice surface and several instruments;

[0027] S02, then, use 200~500ul of chloral hydrate to anesthetize the prepared wild mouse to death by intraperitoneal injection, and immediately use 20ml of phosphate buffered saline solution to perfuse and wash the blo...

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Abstract

The invention discloses a method for preparing a single-cell suspension of aortic cells of a mouse. The method is characterized by specifically comprising the following steps of firstly, preparing 1 ml of an enzyme mixed solution, the surviving healthy wild mouse, an operating table of which the whole tabletop is a flat ice surface and a plurality of instruments; and then, performing intraperitoneal injection anesthesia on the prepared wild mouse by 200-500 microliters of chloral hydrate until the mouse is dead, immediately performing perfusion flushing on blood vessels of the wild mouse by 20ml of a phosphate buffer salt solution, performing overall disinfection on the dead wild mouse by alcohol with the volume fraction of 75%, performing thoracotomy andlaparotomy after disinfection is completed, and exposing the heart and the aorta. The whole operation process is completed on the operating table. Preparation of a single-cell suspension of vascular tissues of the mouse by using a shearing and mixed enzyme combined digestion process is simple, convenient and feasible, more immune cells exist, and an effective tissue single-cell suspension preparation method is provided for research of vascular immune cells.

Description

technical field [0001] The invention relates to the technical field of cell quantitative analysis, in particular to a method for preparing a single-cell suspension of mouse aortic cells. Background technique [0002] Flow cytometry is a technology based on single cells, which is widely used in basic research and clinical practice, such as immunophenotyping of mouse vascular cell membranes, and the preparation of single-cell suspensions of mouse vascular tissue is a method of flow cytometry. prerequisite for cell analysis. The preparation of the existing vascular tissue single cell suspension is relatively difficult, and it is difficult to obtain single cells with high yield, high activity and strong function, which hinders further experimental research, and the number of cells is insufficient, cell debris is too much, and cell adhesion Agglomeration, etc. can lead to the failure of the experiment. Therefore, how to make a single cell suspension with better quality is a pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078
CPCC12N5/0634C12N2509/00
Inventor 赵艳芳
Owner ANHUI NORMAL UNIV
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