A kind of construction method of azoospermia mouse model

A construction method and mouse model technology, which is applied in the field of construction of azoospermia mouse models, can solve the problems of poor stability, low modeling rate, and stability of modeling rate restricting the research progress of non-obstructive azoospermia , to achieve the effect of good stability and high molding rate

Active Publication Date: 2021-04-06
THE SIXTH AFFILIATED HOSPITAL OF SUN YAT SEN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the existing methods are different, they all have the characteristics of low molding rate, poor stability and high mortality rate.
The lack of high modeling rate and good stability construction scheme seriously restricts the research progress of non-obstructive azoospermia

Method used

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  • A kind of construction method of azoospermia mouse model
  • A kind of construction method of azoospermia mouse model
  • A kind of construction method of azoospermia mouse model

Examples

Experimental program
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Effect test

Embodiment 1

[0034] A kind of embodiment of the construction method of azoospermia mouse model of the present invention, comprises the steps:

[0035] (1) C57BL / 6 male mice, which are sexually mature at 8-12 weeks, are used as model animals, with a body weight of 25-30g and no more than 35g; the modeling dose (busulfan) is 40mg / kg, and the cycle is 36 days;

[0036] (2) Taking a mouse with a body weight of 25g as an example, 1mg of busulfan is needed, dissolved in 0.05ml DMSO, prepared for immediate use, diluted to 2ml with normal saline (the ratio of DMSO to the total liquid volume is 2.5%), and transported on ice to animal center;

[0037] (3) After the mice enter the animal center after routine disinfection, shake the diluent well, draw 0.5ml diluent with an insulin needle (refer to Table 1 for details), disinfect the lower abdomen with alcohol, insert the needle after it is slightly dry, and draw back to confirm that it is correct. Complete a single intraperitoneal injection;

[0038...

Embodiment 2

[0042] The modeling effect comparison of the busulfan of embodiment 2 different doses

[0043]Model mice were prepared using the modeling method of Example 1, the difference being the dose of busulfan, and the modeling effects of three different doses of 20 mg / kg, 30 mg / kg, and 40 mg / kg were compared. The result is as figure 1 shown by figure 1 It can be seen that, on the 36th day, in the seminiferous tubules of the mouse testis treated by two doses of 20mg / kg and 30mg / kg, spermatogenic cells were still visible, while in the seminiferous tubules of the mouse testis treated by the dose of 40mg / kg, only the same There were no spermatogenic cells in the basal part of the Sertoli cells, therefore, 40mg / kg dose of busulfan is suitable as a modeling dose.

Embodiment 3

[0044] Example 3 Effects of time on the seminiferous tubules and testis of model mice

[0045] Model mice were prepared using the modeling method in Example 1, the difference being that the time after busulfan injection was different, and the changes in the testes of the mice were compared after 0 day, 9 days, 18 days, 27 days, and 36 days. The result is as figure 2 shown by figure 2 It can be seen that from the 9th day to the 18th day, the seminiferous tubules and the testis / body weight coefficient changed drastically, suggesting that the model may be in a reversible stage before the 18th day, and gradually turn into an irreversible state after the 18th day.

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Abstract

The invention discloses a method for constructing an azoospermia mouse model, which comprises the following steps: (1) providing a sexually mature male mouse with a body weight of 25-35 g; (2) dissolving busulfan in DMSO and diluting it with normal saline (3) Alcohol disinfection of the lower abdomen of the mouse, after drying, draw the diluted solution obtained in step (2) and insert the needle, and after withdrawing to confirm that it is correct, complete the first intraperitoneal injection; after that, inject 3 times at intervals within the same day, After 35-37 days, the mouse model of azoospermia was obtained. The method of the present invention has high molding rate and good stability; according to the present invention, 84 C57BL / 6 mice have been accumulatively completed modeling of azoospermia, and the total number of moldings identified by testis HE staining is 84 / 84, and the molding rate is 84 / 84. 100%; adopt the method of the present invention to build the model, the mortality rate of the mice is 0%; according to the method of the present invention, the total number of deaths of 84 C57BL / 6 mice that have been injected is 0, and the mortality rate is 0%.

Description

technical field [0001] The invention relates to the technical field of animal model construction, in particular to a method for constructing an azoospermia mouse model. Background technique [0002] According to whether the vas deferens is unobstructed, azoospermia can be divided into obstructive azoospermia and non-obstructive azoospermia. Non-obstructive azoospermia is mainly caused by testicular spermatogenesis dysfunction, and clinical treatment is difficult. Currently, there is no effective drug treatment for this part of patients, and surgical sperm retrieval is mainly relied on, such as testicular biopsy, open biopsy or microscopic sperm retrieval. Operation. In recent years, more and more methods have been reported for the treatment of non-obstructive azoospermia, such as spermatogonial stem cell transplantation. With the development of cell replacement therapy for non-obstructive azoospermia, it is necessary to obtain a stable animal model of azoospermia for exper...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/255
CPCA61K9/0019A61K31/255
Inventor 刘贵华邓春华谢云邓存灿杨其运张弛吕林艳姚嘉慧
Owner THE SIXTH AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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