利用白消安制备公鸡生殖前体细胞移植受体的研究

Study on Preparation of Roosters Germ Precursor Cells Transplantation Recipients by Injecting Busulfan

该试验旨在开发适合制备鸡生殖前体细胞移植受体公鸡的方法。试验选用5-7月龄、体重相近、繁殖性能正常的81只岭南黄公鸡进行试验。试验分两次开展,试验一根据公鸡体重计算白消安用量,试验二不以公鸡体重来计算白消安用量,而是每组固定白消安用量。通过分析注射白消安后第15天、30天、45天或60天公鸡的血常规指标、体重、睾丸重、死亡率、曲精细管直径、周长和面积及生育能力,评价白消安直接注射到年轻公鸡睾丸中制备鸡生殖前体细胞移植受体方法的效果。试验一结果表明,注射白消安后第15天仅有1只注射量为1.2 mg/kg的公鸡睾丸部分曲精细管内出现生精细胞减少现象,对血常规指标、死亡率、睾丸大小和重量以及受精率均无影响,而其他鸡只与对照组相比无明显变化。试验二结果表明,公鸡睾丸注射30-50mg/单侧白消安后第15和30天,睾丸大小、重量明显变小,第30天后组织切片分析显示睾丸内精曲细管膜底部和腺腔排空,第45天注射量为30-40mg/单侧的公鸡睾丸内大部分精原干细胞可以恢复,白消安注射组在第15-45天公鸡配种后受精率明显下降,但整个过程中公鸡体重和血常规指标无显著影响。综上表明,公鸡睾丸注射白消安可以清除内源性精原干细胞,且睾丸正常生理生殖功能不受影响,注射剂量为30-40mg/单侧后第30天,公鸡睾丸内精曲小管膜底部和腺腔排空,此时是注射生殖前体细胞最佳时间。

To develop a method suitable for preparing chicken reproductive precursor cell transplant recipient rooster.In this experiment,81 Lingnan yellow roosters with similar body weight and normal reproductive performance were selected for the experiment.The test was carried out in two times.The first test was to calculate the busulfan dosage according to the body weight of the roosters.Direct injection of busulfan into males was evaluated by analyzing blood routine indexes,body weight,testicular weight,mortality,diameter of seminiferous tubules,circumference and area,and fertility of males on day 15,30,45 or 60 after injection of busulfan.Effects of a method for preparing chicken reproductive precursor cell transplant recipients in the testes of young roosters.The results of test 1 showed that only one rooster with an injection dose of 1.2 mg/kg showed decreased spermatogenic cells in the seminiferous tubules of the testis on the 15th day after the injection of busulfan.Size and weight and fertilization rates were not affected,while other birds showed no significant change compared to the control group.The results of test 2 showed that the size and weight of the testis were significantly reduced on the 15th and 30th days after injection of 30 mg/unilateral to 50 mg/unilateral busulfan into the testis of roosters.The tissue section analysis after the 30th day showed that the seminiferous tubule membrane in the testis was obtained.The bottom and gland cavity were emptied,and most spermatogonial stem cells in the testicles of males injected with an injection dose of 30 mg/unilateral to 40 mg/unilateral on the 45th day could be recovered.Fertilization rate decreased significantly,but rooster body weight and blood routine indexes had no significant effect during the whole process.In conclusion,the testicular injection of busulfan can clear endogenous spermatogonial stem cells,and the normal physiological reproductive function of the testis is not affected.The injection dose is 30 mg/unilateral to 40 mg/unilateral on the 30th day after,the bottom of seminiferous tubule membrane and gland cavity in the testis of rooster are emptied,and this time is the best time to inject reproductive progenitor cells.