ABSTRACT
Objective To establish a rapid monitoring method of the three common bacteria in mice frozen resources, such as embryo, sperm, etc. Methods To extract DNA of the three positive bacteria( Staphylococcus auerus, Klebsiella?pneumoniae and β?hemolyticstreptococcus) , and establish PCR monitoring method of the three positive strains through designing primer and refining PCR condition. Then extract total DNA of the frozen resources, detect the DNA according to the PCR condition of the three positive bacteria, some samples were detect by fluorescence quantitative PCR at the same time. Results ①successfully establish a PCR detection method of the three positive bacteria, the minimum detectable concentration of Staphylococcus auerus, Klebsiella pneumoniae and β?hemolytic streptococcus is 4?19 × 10 -5 ng/μL, 1?98 × 10 -5 ng/μL and 1?07 × 10 -3 ng/μL. ②Proved that the three bacteria doesn ’ t exist in the sample by normal PCR and fluorescence quantitative PCR methods. Conclusions Establising a rapid monitoring method of common pathogens of frozen embryo and sperm in mice.
ABSTRACT
Objective To compare the effects of different transplantation sites on the outcome of testicular grafts in mice, and to provide a basis for development and application of this technique in relevant research .Method 5-day old and 4-week old SPF male C57BL/6J mice were used in this study .Three groups of testicular transplantation , i.e.dorsal subcutaneous transplantation ( 5 mice, 40 testes ) , transplantation inside the testicular tunica albuginea ( 6 mice, 12 testes), and were subrenal capsule transplantation (10 mice,15 testes) groups were set up for evaluating the effect of transplantation site on the outcomes in mice .Sham operation (4 mice) and castration (4 mice) groups were also used in this study.The mice were sacrificed at 8 weeks after transplantation and the transplanted testes were collected for analysis of their weight, transplant survival, weight gain, and germ cell differentiation.Results There were significant differences of the testicular germ cell differentiation in different transplantation groups .The germ cell differentiation was best in the in-tra-tunica albuginea transplantation group , and were similar to that in the sham operation group .The germ cell differentia-tion rate was 100%in the intra-tunica albuginea transplantation group , 29.2% in the subcutaneous transplantation group and 0%in the subrenal capsule transplantation group .Conclusions Transplantation beneath the testicular tunica albug-inea is the most favorable site for germ cell differentiation , and dorsal subcutaneous transplantation is an alternative choice . Subrenal capsule transplantation is not appropriate for preservation of male reproductive organs in mice .
ABSTRACT
Objective To compare the effect of three cryoprotectants on vitrification-cryopreservation of C57BL/6J mouse epididymis.Method Epididymises from 6-8-week old male C57BL/6J mice (age) were cryopreserved using DMSO, PROH and R18S3 cryoprotectant solution and thawed , respectively.The morphology of sperm from thawed epididy-mis, the rate of post-thawing survival and reproductive capacity were determined to evaluate the freezing efficiency of the three cryoprotectants .Results The sperms from thawed epididymis of the three groups were all well-preserved structural-ly.The survival rate of sperms was 88.17%, 61.17% and 16.83% in the PROH, DMSO and R18S3 cryoprotectant solu-tions, respectively, and there were significant differences between the three cryopreservation groups (P<0.05 for all). The number of pups from the DMSO , PROH and R18S3 groups were 13, 8 and 17 mice after ICSI and embryo implantation , respectively.Conclusions All the three cryoprotectants DMSO , PROH and R18S3 solutions are suitable for vitrification-cryopreservation of C57BL/6J mouse epididymis.But PROH is the preference of these cryoprotectant solutions .