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A kind of method and application of muskrat scent gland secretory cell isolated and cultivated in vitro

A technology for the separation and cultivation of muskrat scent glands, which is applied in the field of cell biology, can solve the problems of time-consuming, financial and manpower consumption, and low yield and quality of secretory cells, and achieve the effect of shortening the process of separation and purification and simplifying the steps of separation and cultivation in vitro

Inactive Publication Date: 2018-10-09
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the yield and quality of secretory cells obtained by the method of mixed culture after protease digestion are not high, and it is a great waste of time, financial resources and manpower

Method used

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  • A kind of method and application of muskrat scent gland secretory cell isolated and cultivated in vitro
  • A kind of method and application of muskrat scent gland secretory cell isolated and cultivated in vitro
  • A kind of method and application of muskrat scent gland secretory cell isolated and cultivated in vitro

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1) Take out the scent gland tissue of the 6-month-old muskrat in the fragrance-producing stage for the test, and quickly place it in 500 U / ml penicillin and streptomycin PBS pre-cooled at 4°C, wash it for 5 times, and use it for later use ;

[0034] 2) Remove the residual fat, blood vessels and connective tissue around the scent gland tissue, peel off the peripheral film of the scent gland tissue, and cut the scent gland tissue into uniform small pieces of tissue with a thickness of 0.7cm with an animal tissue cutting instrument;

[0035] 3) Use 4°C pre-cooled 500U / ml penicillin and streptomycin PBS to wash even small pieces of fragrant gland tissue with a thickness of 0.7cm. After washing 8 times, observe the supernatant from turbid to clear, and centrifuge at 1000rpm / min for 5min , discard the supernatant;

[0036] 4) Add 5ml of 150U type II collagenase to a 15ml centrifuge tube containing a small piece of fragrant gland tissue, pipette evenly, then quickly place it ...

Embodiment 2

[0042] The difference with embodiment 1 is:

[0043]The scent glands were cut into 0.8 cm thick scent gland tissue fragments with an animal tissue cutter, the concentration of type 2 collagenase was 200 U, and the volume of adding type 2 collagenase was 6 ml. The cells obtained after culturing were not significantly different from the cells obtained in Example 1 upon observation.

Embodiment 3

[0045] This embodiment provides a method for culturing muskrat scent gland tissue cells commonly used in the prior art, and the specific steps are as follows:

[0046] 1) The scent gland tissue of a one-year-old male muskrat in the fragrance-producing stage was taken out in a sterile environment, washed with 4°C pre-cooled penicillin and streptomycin PBS for 5 times, and the residual fat around the scent gland tissue was removed. Blood vessels and connective tissue, peel off the peripheral film of the scent gland tissue, use ophthalmic scissors or a scalpel to cut the scent gland tissue into a uniform thickness of 1mm 3 Small pieces of tissue;

[0047] 2) Use 4°C pre-cooled penicillin and streptomycin PBS to wash small pieces of scented gland tissue, wash 8 times, observe that the supernatant turns from turbid to clear, centrifuge at 1000rpm / min for 5min, and discard the supernatant;

[0048] 3) Add 6ml 150U type II collagenase to a 15ml centrifuge tube containing a small pie...

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Abstract

The invention discloses a method for separating and culturing muskrat scent gland secretory cells in vitro and its application, and belongs to the technical field of cell biology. In the present invention, the scent gland tissues of muskrats in the fragrance secretion stage are taken, and immediately placed in a 4°C pre-cooled phosphate buffer solution containing penicillin and streptomycin for cleaning treatment, and the peripheral tissues of the scent glands are removed. The scented gland tissue was washed with a 4°C pre-cooled phosphate buffer solution containing penicillin and streptomycin, then centrifuged, digested with type II collagenase, centrifuged after digestion, and the precipitated tissue was cultured and separated. The invention simplifies the in vitro separation and culture steps of the muskrat scent gland secreting cells, only needs to obtain uniform small pieces of scent gland tissue, and can directly carry out the culture operation, and can ensure that the growth condition of the cells is close to the microenvironment in the body. At the same time, the present invention can simply and quickly obtain secretory cells with high purity, greatly shortening the process of separating and purifying secretory cells.

Description

technical field [0001] The invention relates to a method for separating and culturing muskrat scent gland secretory cells in vitro and its application, belonging to the technical field of cell biology. Background technique [0002] The musk secreted by muskrats is also called muskrat scent, which contains the same main components as natural musk, such as muskone, normusketone, alkanone, etc., and can be widely used in the fields of traditional Chinese medicine and spices. Muskrat incense is secreted by the secretory cells of the muskrat scent glands, so the isolation and culture of muskrat scent gland secretory cells in vitro can not only provide experimental materials for the study of the secretion mechanism of the scent glands, but also provide a theoretical basis for the application of in vitro production of muskrat scent. [0003] At present, most of the techniques for isolating and culturing muskrat scent gland secretory cells in vitro adopt the method of mixed culture ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
Inventor 李春义王文英安培培郭倩倩
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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