Preparation method of gastric mucosa tissue protein samples

A technology for tissue protein and gastric mucosa, applied in the field of biomedicine, can solve the problems of cumbersome process and long time, and achieve the effect of solving cumbersome process, good biological repeatability and good protein separation effect.

Inactive Publication Date: 2019-04-02
ZHEJIANG CHINESE MEDICAL UNIVERSITY
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Problems solved by technology

[0006] In summary, the existing problems in the prior art are: the existing g

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  • Preparation method of gastric mucosa tissue protein samples

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[0017] In order to make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention, but not to limit the present invention.

[0018] Such as figure 1 As shown, the method for preparing a gastric mucosal tissue protein sample provided by the embodiment of the present invention includes the following steps:

[0019] S101: Take out the stomach tissue of the fetus within 12 hours of birth, wash the surface of the stomach tissue twice with sterile PBS, remove the connective tissue, and then wash it again with sterile PBS;

[0020] S102: Cut the stomach pouch longitudinally, rinse three times with sterile PBS, rinse the contents and mucus of the stomach; cut the washed stomach tissue into 2-3mm 2 Then transfer to a 15ml centrifuge tube, add a mixed en...

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Abstract

The invention belongs to the technical field of biomedical science, and discloses a preparation method of gastric mucosa tissue protein samples. The preparation method comprises the following steps oftaking stomach tissue from fetal rats born for 12h or below, cleaning stomach tissue surfaces twice with bacteria-free PBS, removing reticular tissue, and performing cleaning once with the bacteria-free PBS; cutting the thoroughly-washed stomach tissue to obtain broken blocks of 2-3mm<2>, transferring the broken blocks into a centrifugal tube of 15ml, adding mixed enzymes of 0.1% of collagenase II and 0.2mg/ml of hyaluronidase, and performing vibration with a water bath for digestion; and finally, re-suspending cells with a complete culture medium, transferring the cells to a culture bottle,placing the culture bottle under the condition of an incubator of 37 DEG C and 5% CO2, and performing culturing. According to the preparation method, gastric mucus can be effectively removed, the gastric mucus is prepared into a unicellular suspension, unicellular protein analysis and study of a gastric mucosa flow cytometry and the like can be performed, and preparation is provided for development of cell components in stomach tissue. The problems of tedious process, long time consumption and high expense can be solved.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for preparing a gastric mucosa tissue protein sample. Background technique [0002] At present, the existing technologies commonly used in the industry are as follows: [0003] Fresh gastric tissue specimens were taken during the operation, and tissue pieces about 2.0cm×2.0cm×0.5cm in size were taken, immediately placed in ice phosphate buffered saline (PBS), and processed into single-cell suspensions according to the following steps within 4 hours: Rinse with normal saline, put 30-50ml of 0.1% pepsin solution (prepared with 0.1N HCL), and put it in an environment of 37°C for about 30 minutes. After taking it out, use tweezers or a thin piece (such as a cell scraper) to remove the digested flocculent The mucus is removed from the gastric mucosa, rinsed with PBS, immediately placed in 1.2-2.4u / ml dispase (prepared in PBS), in an environment of 37°C, with...

Claims

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Application Information

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IPC IPC(8): C12P21/06
CPCC12P21/06
Inventor 郑小伟孔丽娅余王琴刘晓谷
Owner ZHEJIANG CHINESE MEDICAL UNIVERSITY
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