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.
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[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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