SD rat thoracic artery smooth muscle cell separation and culture method

Abstract

The invention belongs to the technical field of biology, and specifically discloses an SD thoracic rat artery smooth muscle cell separation and culture method. The method comprises the following steps: (1) cutting SD rat thoracic artery in a sterile condition; (2) cutting the thoracic artery into long sections with a length of 3 to 5 centimeters, placing the sections in a PBS solution, cutting the thoracic artery in the vertical direction, removing the fat and connective tissues around the blood vessels, then soaking thoracic artery in collagenase (II); (3) taking out processed thoracic artery, and removing the external membrane; (4) incubating the thoracic artery in digestive juice, and then transferring thoracic artery to a DMEM culture medium; (5) transferring thoracic artery to a centrifuge, carrying out centrifugation, collecting the precipitate (namely thoracic artery smooth muscle cells), then placing the obtained thoracic artery smooth muscle cells into a fetal calf serum DMEM culture bottle, then transferring thoracic artery smooth muscle cells to an incubator, culturing the thoracic artery smooth muscle cells in the incubator, and then carrying out subculture. The provided method can greatly increase the number of obtained cells and improves the material utilization rate. Furthermore, the operation is simple and convenient.

Description

technical field

[0001] The invention belongs to the field of biotechnology, in particular to a method for separating and culturing SD rat thoracic aortic smooth muscle cells. Background technique

[0002] Arterial smooth muscle cells have a variety of physiological functions, can produce and secrete many biologically active substances, and play an important role in maintaining vasoconstriction and anticoagulation. The application and development of this method provides an important research method for the study of the pathogenesis and prevention of certain cardiovascular diseases. However, the existing separation methods are immature, not only the number of isolated cells is small, but also the material utilization is low, and the operation is troublesome. Contents of the invention

[0003] The purpose of the present invention is to provide a method for separating and culturing SD rat thoracic aortic smooth muscle cells, which not only greatly increases the number of isol...

Examples

Embodiment 1

[0031] A method for separating and culturing SD rat thoracic aortic smooth muscle cells, comprising the following steps:

[0032] 1) Select SD rats with a weight of 100-150g, cut out the thoracic aorta of SD rats under sterile conditions and remove blood stains, and set aside;

[0033] 2) Cut the thoracic aorta into a 3-5cm long section and put it in PBS solution, then cut the thoracic aorta longitudinally, remove the fat and connective tissue outside the blood vessel, and then put the thoracic aorta into the mass Collagenase at a concentration of 2 g / L Soak in medium for 20 min, set aside;

[0034] 3) Take out the thoracic aorta treated in step 2), remove the adventitia on the thoracic aorta, and set aside;

[0035] 4) Put the thoracic aorta treated in step 3) into the digestive solution, and transfer to a temperature of 37 ℃, C0 2 Incubate in an incubator with a volume concentration of 5% for 2 h, and then put it into a DMEM medium with a concentration of 20% fetal bovin...

Embodiment 2

[0043] A method for separating and culturing SD rat thoracic aortic smooth muscle cells, comprising the following steps:

[0044] 1) Select SD rats with a weight of 100-150g, cut out the thoracic aorta of SD rats under sterile conditions and remove blood stains, and set aside;

[0045] 2) Cut the thoracic aorta into a 3-5cm long section and put it in PBS solution, then cut the thoracic aorta longitudinally, remove the fat and connective tissue outside the blood vessel, and then put the thoracic aorta into the mass Collagenase at a concentration of 2 g / L Soak in medium for 30 minutes, set aside;

[0046] 3) Take out the thoracic aorta treated in step 2), remove the adventitia on the thoracic aorta, and set aside;

[0047]4) Put the thoracic aorta treated in step 3) into the digestive solution, and transfer to a temperature of 37 ℃, C0 2 Incubate in an incubator with a volume concentration of 5% for 2 h, and then put it into a DMEM medium with a concentration of 20% fetal bo...

technical field

[0001] The invention belongs to the field of biotechnology, in particular to a method for separating and culturing SD rat thoracic aortic smooth muscle cells. Background technique

[0002] Arterial smooth muscle cells have a variety of physiological functions, can produce and secrete many biologically active substances, and play an important role in maintaining vasoconstriction and anticoagulation. The application and development of this method provides an important research method for the study of the pathogenesis and prevention of certain cardiovascular diseases. However, the existing separation methods are immature, not only the number of isolated cells is small, but also the material utilization is low, and the operation is troublesome. Contents of the invention

[0003] The purpose of the present invention is to provide a method for separating and culturing SD rat thoracic aortic smooth muscle cells, which not only greatly increases the number of isol...

Examples

Embodiment 1

[0031] A method for separating and culturing SD rat thoracic aortic smooth muscle cells, comprising the following steps:

[0032] 1) Select SD rats with a weight of 100-150g, cut out the thoracic aorta of SD rats under sterile conditions and remove blood stains, and set aside;

[0033] 2) Cut the thoracic aorta into a 3-5cm long section and put it in PBS solution, then cut the thoracic aorta longitudinally, remove the fat and connective tissue outside the blood vessel, and then put the thoracic aorta into the mass Collagenase at a concentration of 2 g / L Soak in medium for 20 min, set aside;

[0034] 3) Take out the thoracic aorta treated in step 2), remove the adventitia on the thoracic aorta, and set aside;

[0035] 4) Put the thoracic aorta treated in step 3) into the digestive solution, and transfer to a temperature of 37 ℃, C0 2 Incubate in an incubator with a volume concentration of 5% for 2 h, and then put it into a DMEM medium with a concentration of 20% fetal bovin...

Embodiment 2

[0043] A method for separating and culturing SD rat thoracic aortic smooth muscle cells, comprising the following steps:

[0044] 1) Select SD rats with a weight of 100-150g, cut out the thoracic aorta of SD rats under sterile conditions and remove blood stains, and set aside;

[0045] 2) Cut the thoracic aorta into a 3-5cm long section and put it in PBS solution, then cut the thoracic aorta longitudinally, remove the fat and connective tissue outside the blood vessel, and then put the thoracic aorta into the mass Collagenase at a concentration of 2 g / L Soak in medium for 30 minutes, set aside;

[0046] 3) Take out the thoracic aorta treated in step 2), remove the adventitia on the thoracic aorta, and set aside;

[0047]4) Put the thoracic aorta treated in step 3) into the digestive solution, and transfer to a temperature of 37 ℃, C0 2 Incubate in an incubator with a volume concentration of 5% for 2 h, and then put it into a DMEM medium with a concentration of 20% fetal bo...

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