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Preparation method of aortic cell single-cell suspension

A single cell suspension and cell suspension technology, applied in the field of cell biology, can solve the problems of low efficiency and poor cell quality, and achieve the effect of high single cell yield, high cell activity, and poor cell quality.

Pending Publication Date: 2022-06-07
THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV
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Problems solved by technology

[0004] Aiming at the deficiencies in the prior art, the present invention provides a method for preparing aortic cell single-cell suspension to solve the technical problem of poor cell quality obtained from the low efficiency of traditional aortic cell single-cell suspension in the related art

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  • Preparation method of aortic cell single-cell suspension
  • Preparation method of aortic cell single-cell suspension
  • Preparation method of aortic cell single-cell suspension

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preparation example Construction

[0028] The invention provides a preparation method of aortic cell single cell suspension, comprising:

[0029] S1, take the aortic vascular tissue, wash it, and then aseptically culture it in a 1640 medium containing 8% to 20% bovine serum at the first temperature, for subsequent use;

[0030] S2. Take the spare aortic vascular tissue in step S1, peel off the fat and connective tissue, and chop it to 1-2 mm 3 tissue fragments, spare;

[0031] S3. Take the spare tissue fragments and the prepared tissue digestion suspension in step S2 at a ratio of 1:(5-8) by volume, mix them and place them in a 37°C environment for the first digestion reaction. For a period of time, until there are no formed tissue fragments, filter, centrifuge, and remove the supernatant to obtain the first cell suspension;

[0032] S4. Take the first cell suspension and trypsin obtained in step S3 at a ratio of 1:(1.8-3) by volume, mix them, and place them in a 37°C environment to carry out the second diges...

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Abstract

The invention provides a preparation method of a single-cell suspension of aortic cells, which comprises the following steps: firstly treating aortic vascular tissues, and then carrying out secondary digestion reaction to obtain the single-cell suspension with high single cell yield, large quantity, high cell activity and strong cell function. The technical problem that a traditional aortic cell single-cell suspension is low in efficiency, and obtained cells are poor in quality is solved.

Description

technical field [0001] The invention relates to the technical field of cell biology, in particular to a preparation method of aortic cell single cell suspension. Background technique [0002] Aortic disease is a relatively dangerous cardiovascular disease. Common aortic dissection, intramural hematoma, atherosclerosis, aneurysm, Marfan syndrome, congenital aortic disease, traumatic disease, aortic inflammation, etc. The mechanism of aortic disease is complex, and there are many types of cells involved. With the development of single-cell sequencing technology, the analysis results of relevant participating cells have an important impact on the conquest of aortic disease. However, due to the high toughness of the aortic blood vessels, the preparation of single cell suspension is difficult, and it is difficult to obtain single cells with high efficiency, high activity and strong function for the study of related pathology. [0003] Therefore, the emergence of a preparation m...

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/069C12N2509/10C12N2509/00
Inventor 贺文辉李军程伟余三九何萍许志臻燕朝均钟丹戴双双
Owner THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV
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