Separation and passage method of pregnant rat placenta vascular smooth muscle cell

A technique of vascular smooth muscle and separation method, applied in cell dissociation method, vascular endothelial cells, embryonic cells, etc., can solve the problem of difficult separation of blood vessels of pregnant mouse placenta, reduce the risk of cell contamination, simple and efficient extraction, reduce Effects of time of tissue exposure to air

Inactive Publication Date: 2017-08-18
CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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Problems solved by technology

[0004] In order to solve the difficult problem of separating blood vessels from the placenta of pregnant mice, the object of the present invention is to provide a method for isolating and subcultureing placental vascular smooth muscle cells of pregnant mice, a more convenient and efficient method for extracting placental vascular smooth muscle cells

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  • Separation and passage method of pregnant rat placenta vascular smooth muscle cell
  • Separation and passage method of pregnant rat placenta vascular smooth muscle cell
  • Separation and passage method of pregnant rat placenta vascular smooth muscle cell

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Embodiment

[0033] This embodiment provides a method for isolating vascular smooth muscle cells of pregnant mouse placenta, the schematic diagram of which is as follows figure 1 As shown, it includes the following steps:

[0034] After the pregnant mouse was anesthetized (50 mg / kg pentobarbital sodium), it was sprayed with 75% alcohol for disinfection, and after laparotomy, the uterus was separated rapidly and placed in precooled sterile PBS solution;

[0035] Cut the uterus in an ultra-clean bench, peel off the placenta from the uterus, cut off the junction between the fetus and the umbilical cord, put the separated placenta with the umbilical cord in pre-cooled sterile PBS, and place the tissue in a 4°C refrigerator or store on ice;

[0036] Put 1 placenta into a black plastic dish containing pre-cooled sterile PBS solution on ice, connect a 30G 1 / 2IN sterile injection needle to a 5mL (or above capacity) sterile syringe barrel , draw 3mL of pre-cooled sterile PBS solution, under a bod...

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Abstract

The invention provides a separation and passage method of a pregnant rat placenta vascular smooth muscle cell. The method comprises the steps of taking a pregnant rat placenta, conducting pretreatment, afterwards injecting a mixed solution of a serum-free DMEM F12 culture medium containing agarose, ferroferric oxide and penicillin streptomycin, cutting the placenta into pieces after the mixture is congulated, adsorbing placenta tissue containing iron under the effect of magnetic force, using a collagenase II solution to conduct resuspending, after enzymatic digestion is completed, adding a DMEM F12 total culture medium to passivate collagenase II, and after washing is conducted, adding the DMEM F12 total culture medium again to conduct resuspending, wherein obtained tissue blocks grow sticking to the wall, and then the vascular smooth muscle cell is grown so that subculture can be conducted. According to the separation and passage method of the pregnant rat placenta vascular smooth muscle cell, compared with a traditional method, the placenta vascular smooth muscle cell can be conveniently and efficiently extracted, in the whole process, blood vessels do not need to be dissociated out, the operation is simpler, the time of the tissue for being exposed is shortened, and the risk of cell contamination is lowered.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for separating and passing on placental vascular smooth muscle cells of pregnant mice. Background technique [0002] Pregnant mouse placental vascular smooth muscle cells are isolated from the placental arterial vessels of pregnant mice in the third trimester and can be cultured in vitro. Because human placenta is not easy to obtain and has large individual differences, mice are commonly used as animal models, and their specimens are easy to obtain and have great impact on human placenta. The response to drugs and other stimuli is highly similar to that of the human body, which can simulate the human body very well. Therefore, the placental vascular smooth muscle cells of pregnant mice are an important tool for studying the medical basis of placental vascular diseases. [0003] However, there is no mature method to isolate and culture vascular smooth muscle cells from the plac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077C12N5/073
CPCC12N5/0605C12N5/0691C12N2509/00
Inventor 刘慧荣贺春雨张苏丽王美丽白丽娜王鹏丽
Owner CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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