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Preparation method of spermatogonial stem cell transplantation receptor

A technology for spermatogonial stem cells and transplant recipients, which is applied in the field of spermatogonial stem cell transplant recipient preparation, can solve the problems of insignificant effects and affect the results of transplantation, and achieve the effects of avoiding death, reducing injury, and avoiding the death of recipient animals.

Active Publication Date: 2012-12-19
洛阳中科基因检测诊断中心有限公司
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  • Claims
  • Application Information

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Problems solved by technology

However, the testicular microenvironment of the recipients treated with this method recovers quickly, and the highest transplantation efficiency can only be obtained within the best tolerance period, so the recipients prepared by this method must strictly pay attention to the transplantation time, otherwise the transplantation results will be affected
Although the heat treatment method has no toxic effect on animals, its effect is relatively inconspicuous, and the time requirement is strict, so it is seldom used.

Method used

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  • Preparation method of spermatogonial stem cell transplantation receptor
  • Preparation method of spermatogonial stem cell transplantation receptor
  • Preparation method of spermatogonial stem cell transplantation receptor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1 receptor preparation

[0029] Recipient: 1-2 month old CD1 (ICR) strain normal breeding male mice. purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.

[0030] Busulfan configuration: Dissolve busulfan in DMSO at a concentration of 8 mg / ml, add an equal volume of ultrapure water 0.5 hours before injection, set the final concentration at 4 mg / ml, and bathe in a 37°C water bath to prevent coagulation.

[0031] Injection: Connect a disposable intravenous infusion needle of 0.45mm×15mm to the syringe for standby, the injection concentration is 4mg / ml, the doses are 2mg / kg, 3mg / kg, 4mg / kg, 5mg / kg, and the injection volume is intraperitoneal injection 1 / 10. At the same time, the intraperitoneal injection control group and the blank control group were set up, and the intraperitoneal injection group was injected in the conventional way, and the injection dose was 40 mg / kg.

[0032] Slices: The receptors in the testicular injection group...

Embodiment 2

[0037] Example 2 Sperm stem cell transplantation

[0038] Donor: C57 red fluorescent male mice aged 4-6 days. Purchased from the Genetic Center, Institute of Experimental Animals, Chinese Academy of Medical Sciences

[0039] Donor Cell Suspension Preparation

[0040]Cell suspensions were prepared using a two-step enzymatic digestion method. The C57 red fluorescent mice aged 4-6 days after birth were taken out and sacrificed, and the testes were taken out and quickly placed in calcium-magnesium-free PBS. Take 10-20 testes for each test, remove the tunica albuginea, divide and spread out the seminiferous tubules, remove the visible cell ribbons between the tubules, put them into a centrifuge tube and cut them into pieces with scissors. Add 1ml of collagenase (1mg / ml) and DNase (0.4mg / ml) to digest for 5min, continue pipetting, and centrifuge at 1000r / min for 3min. Discard the supernatant, add 1ml PBS to wash, mix well and centrifuge for 3-4min, repeat 2 times. Discard the s...

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Abstract

The invention discloses a preparation method of a spermatogonial stem cell transplantation receptor. The spermatogonial stem cell transplantation receptor is prepared by the method through testicular busulfan injection. By applying the method disclosed by the invention, the spermatogonial stem cell transplantation receptor can be successfully and quickly prepared, and the larger damages to an animal bone marrow and a hemopoietic system through a celiac busulfan injection method are reduced, so that the animal death is reduced or avoided, and the consumption of drugs can also be reduced, and the cost is reduced. The invention opens up a novel preparation method of the spermatogonial stem cell transplantation receptor. The method does not involve in any special instrument and can be finished only by utilizing the existing conventional instrument and a chemical reagent. The invention can be implemented by ordinary technical personnel in the field. The preparation method of the spermatogonial stem cell transplantation receptor has the advantages of low cost, simplicity and convenience in operation, high efficiency, quickness, safety and feasibility and is beneficial to popularizing, applying, optimizing and innovating of a spermatogonial stem cell transplantation technology.

Description

technical field [0001] The invention belongs to the field of applied biotechnology, and is a novel, high-efficiency, and low-toxic preparation method for recipients of spermatogonial stem cell transplantation, which can be used in spermatogonial stem cell transplantation, preparation of transgenic animals by using spermatogonial stem cells, and protection of endangered species by using spermatogonial stem cells applications in other fields. Background technique [0002] Spermatogonia stem cells are a group of diploid cells near the basement membrane of the seminiferous epithelium with high self-renewal and multilineage differentiation potential. They are the only adult stem cells that can transmit genetic information to the next generation in adult animals. Spermatogonia stem cells can not only continuously self-renew to maintain a constant number of their own population, but also continuously differentiate to produce spermatogonia at all levels, and finally generate sperm. ...

Claims

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Application Information

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IPC IPC(8): A61D7/00
Inventor 王栋朱化彬刘玲王琛郝海生秦彤赵学明
Owner 洛阳中科基因检测诊断中心有限公司
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