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Isolation and culture method for primary rat aortic smooth muscle cells

A technique for the isolation and cultivation of smooth muscle cells, applied in the field of isolation and cultivation of primary rat aortic smooth muscle cells, can solve the problems of impure cell types, poor cell separation, and reduced activity, so as to improve cell activity and yield, improve High adherence rate and high cell viability

Inactive Publication Date: 2016-08-31
王晓冰 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages are that, firstly, the culture time of the primary cells in the tissue block culture method is long, and not every block has cells to germinate, so the number of cell passages and the number of finally obtained cells are small; secondly, the cell types are not pure, and simply use Digestion and isolation of tissue blocks resulted in the incorporation of many other cells other than aortic smooth muscle cells; thirdly, the use of a single collagenase to digest cells resulted in more pipetting during the separation process, poor cell separation and reduced activity; finally, the separation of cells and It is easy to contaminate during the process of culturing cells, resulting in low cell viability and activity

Method used

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  • Isolation and culture method for primary rat aortic smooth muscle cells
  • Isolation and culture method for primary rat aortic smooth muscle cells
  • Isolation and culture method for primary rat aortic smooth muscle cells

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Embodiment 1

[0046] In this example, newborn SD rats were used as the source of cells. First, they were sacrificed by cervical dislocation, soaked in alcohol for 5 minutes, and under a dissecting microscope, the thorax was opened, and the lung tissue was cut off to fully expose the back wall of the thorax. Dissect the aorta starting from the ascending aorta of the heart. Use one tweezer to clamp the ascending aorta, and the other tweezer to gradually peel off the thoracic aorta along the posterior wall of the thoracic cavity until the tail end of the thoracic aorta, that is, before entering the abdominal cavity, and cut off the aorta. After washing the isolated rat aorta with 37°C preheated D-Hanks solution containing 2% bis-antibody, carefully peel off the intima of the blood vessel, and use a small syringe to connect a thin hose (dwelling needle hose) for perfusion37 Preheated perfusate at ℃, the perfusion rate is 5mL / min, the time is 5 minutes, the preperfusate formula is 15mM / L HEPES, ...

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Abstract

The invention provides an isolation and culture method for primary rat aortic smooth muscle cells. With the isolation and culture method provided by the invention, the acquired cells are great in gross quantity and high in both isolation degree and motility rate; the probability of contamination by bacteria, fungi and other cells is low; and the method can realize subculturing, so a good cell culture foundation is laid for related research on blood vessel diseases.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for separating and culturing primary rat aortic smooth muscle cells. Background technique [0002] A major factor in the development and progression of vascular disease is the transformation of vascular smooth muscle cells into a reproductively competent phenotype. Recent studies have shown that smooth muscle cells express calcium channels, ICAM-1 and VCAM-1. Among them, the expression of ICAM-1 and VCAM-1 may be the cause of vascular wall inflammation and further cause vascular diseases. Therefore, the in vitro culture and study of vascular smooth muscle cells can be used to discover and determine new targeted therapies for vascular diseases. [0003] At present, a large number of researchers at home and abroad have carried out studies on the culture and pharmacological characteristics of mouse / rat aortic smooth muscle cells. The core of primary cell culture is t...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0691C12N2509/10
Inventor 王晓冰杨国峰
Owner 王晓冰
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