生物工程技术在SPF级大小鼠生物净化上的应用及其临床价值

Application and Clinical Value of Bioengineering Technology in Biological Purification of SPF Mice

目的:探究在大小鼠生物净化中应用生物工程技术的效果。方法:自2016年1月开始,本研究中心选择常用的C57BL/6J品系小鼠作为研究A组、选择SD大鼠作为研究B组,在繁殖中应用生物工程技术;另选择一部分小鼠与大鼠作为对照A、B组,在其繁殖过程中不做任何干涉随机。每组研究对象15只。实验开始前,对所有鼠的病原微生物进行检测。应用体外受精与胚胎移植的生物工程技术,在HTF培养液中完成受精并发育至2-细胞时期,将胚胎移植入购买来并激素处理过的代孕SPF级ICR雌鼠的输卵管中。代孕ICR雌鼠生下子代,子代3周离乳后,对代孕ICR雌鼠和随机挑选的子代一只进行检测。统计研究A、B组的采集卵子数、2-细胞数与移植胚胎数。统计四组研究对象的平均每胎产仔数、存活率。统计净化前后四组研究对象的病原微生物检出率及净化率。结果:研究A、B组生物工程技术指标(采集卵子数、2-细胞数与移植胚胎数)比较,差异均无统计学意义(P>0.05);四组研究对象平均每胎产仔数与存活率比较,差异均无统计学意义(P>0.05);四组研究对象在合笼与体外受精前,均尾静脉取血进行血霉菌培养。仔鼠出生3周离乳后,每胎随机选一只与其代孕ICR母鼠一起进行尾静脉取血,进行血霉菌培养,对照A、B组的净化率均显著低于研究A、B组(P<0.05)。结论:在小鼠大鼠的生物净化中应用体外受精与胚胎移植的生物工程技术,在不影响代孕母鼠的生殖能力与子代存活率的情况下,显著地提升了净化率,达到了研究的目的,从而能为临床动物实验提供更多优质可用的实验动物。

Objective：To explore the application of bioengineering technology in biological purification of large and small mice.Method：Since January 2016,the research center selected the common C57BL/6J strain mice as the study group A and SD rats as the study group B,applied bioengineering techniques to the reproduction, and selected some mice and rats as the control A and B groups,and did not interfere random in their reproduction process.There were 15 subjects in each group.The pathogenic microorganisms of all rats were detected before the experiment.The bioengineering techniques used in vitro fertilization and embryo transfer were used to fertilize and develop into the 2-cells period in the HTF culture.The embryos were transplanted into the oviduct of the SPF grade ICR female rat of the surrogate, which was purchased and treated with hormone.The surrogate ICR female rats were born in the next generation.After three weeks of subgeneration,the surrogate ICR female rats and one offspring were randomly selected.The number of ovum collected,the number of 2-cells and the number of transplanted embryos in the study A and B groups were statistically analyzed.The average number of litter per litter and survival rate of the four groups were analyzed.The detection rate and purification rate of pathogenic microorganisms in four groups before and after statistical purification were statistically analyzed.Result：The biological engineering technical indexes（the number of egg collection,the number of 2-cells and the number of transplanted embryos） of the study A and B groups,there were no statistical significance（P〉0.05）.The average number of births and survival rates and the survival rate of the four groups,there were no significant differences（P〉0.05）.In the four groups,blood samples were taken from the tail vein before the cage and in vitro fertilization.After three weeks of birth,one of the offspring was randomly selected to take the tail vein with a surrogate ICR mouse to take blood from the tail vein to carry out the blood mold culture.The purification rate of the control A and B groups were significantly lower than those of the study A and B groups（P〈0.05）.Conclusion：The bioengineering techniques for the application of in vitro fertilization and embryo transfer in the biological purification of the mice,without affecting the reproductive and progeny survival rate of the surrogate mouse,significantly improve the purification rate and reached the aim of the study,thus providing more quality and available experimental animals for clinical animal experiments.