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Primary mouse thoracic aortic endothelial cell separation and culture method

A technology of thoracic aorta and vascular endothelium is applied in the field of separation and culture of primary mouse thoracic aortic vascular endothelial cells, which can solve the problems of difficult culture and low yield, and achieve the effects of low technical difficulty, eliminating pollution and improving purity.

Inactive Publication Date: 2017-04-26
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] The invention solves the defects and deficiencies in the process of primary isolation and culture of VECs from the aorta in the prior art, and provides a method for the isolation and culture of primary mouse thoracic aortic endothelial cells, which overcomes the existing In the technology, primary VECs are difficult to culture in vitro, and the yield is low, so that the primary cultured VECs have a high proliferation rate

Method used

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  • Primary mouse thoracic aortic endothelial cell separation and culture method
  • Primary mouse thoracic aortic endothelial cell separation and culture method
  • Primary mouse thoracic aortic endothelial cell separation and culture method

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Embodiment

[0054] A method for isolating and culturing primary mouse thoracic aortic endothelial cells, specifically as follows:

[0055] 1. Matrigel pretreatment of the bottom of the orifice plate

[0056] One day before the first ring implantation, the whole bottle of Matrigel Matrigel (purchased from BD Company, USA) was embedded in ice in advance and placed at 4°C overnight to make it fully change from jelly to solution. Under a sterile environment, pipette and mix the product with a cold gun tip, then divide into small tubes (500 μL / tube), keep one tube on ice for later use, and store the rest at -20°C.

[0057] In this embodiment, a 12-well plate is taken as an example, and the melted Matrigel is added into the wells (200 μL / well), and the orifice plate is tilted so that the Matrigel evenly covers the bottom of the wells. In view of the amount of samples taken from two mice, only the two central wells of the 12-well plate were pretreated, and sterile PBS (1 mL / well) was added to t...

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Abstract

The invention provides a primary mouse thoracic aortic endothelial cell separation and culture method, which solves the problems of great difficulty in primary VECs (vascular endothelial cell) in-vitro culture, low cell activity and little output in the prior art. The method comprises: (Step 1) a mouse thoracic aortic section is obtained, and after separation and purification, the mouse thoracic aortic section is produced into an aortic ring; (Step 2) a matrigel is used for pretreating the bottom of a culture vessel; (Step 3) the aortic ring is inoculated to the surface of the matrigel, an endothelial cell culture medium which contains an endothelial cell growth-assisting factor and heparin is then added into the culture vessel for culture; (Step 4) after 3 to 7 days of culture, the aortic ring is removed, and culture is continued. The method is easy and convenient to implement, economical and applicable, and has the advantages of decreasing the probability of impurity cell contamination, increasing the survival rate and output of cells, enabling VECs to have high activity and high multiplication rate, etc..

Description

technical field [0001] The invention belongs to the field of material collection, separation, purification and cultivation of vascular endothelial cells, and relates to a method for separating and culturing primary mouse thoracic aortic vascular endothelial cells. Background technique [0002] Blood vessels are the conduits for the organism to transport blood, provide the necessary oxygen and nutrients for the normal metabolism and growth of the body, and transport away harmful substances. Vascular endothelial cells (VECs), as a layer of multifunctional cells lining the inner wall of blood vessels, have very active endocrine and metabolic functions, and play an extremely important role in the vascular permeability barrier, immune defense and inflammatory response. It can be described as a key factor in maintaining the normal function of blood vessels. However, in some diseases such as tumors, cardiovascular diseases, and inflammatory response diseases, VECs have become impo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/069C12N2501/10C12N2501/91C12N2509/00C12N2513/00
Inventor 甘雪琦张玲
Owner SICHUAN UNIV
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