期刊
REPRODUCTION FERTILITY AND DEVELOPMENT
卷 31, 期 3, 页码 538-546出版社
CSIRO PUBLISHING
DOI: 10.1071/RD18191
关键词
chemical depletion; gamete biology; spermatogenesis; stem cells
资金
- H. H. Shaikh Mohammed bin Rashid Al Maktoum
- United Arab Emirates (UAE)
- Emirate of Dubai
The object of this study was to investigate if testis germ cell transplantation (TGCT) into a heterologous recipient would result in donor-origin spermatogenesis in the dromedary camel. First, we investigated a workable protocol for TGCT in camels, including donor cell isolation, enrichment by density gradient centrifugation (Percoll and Bovicoll), rete testis injection and microsatellite detection of donor and recipient genotypes. Second, the effects of three doses of Dolichos biflorus agglutinin (DBA), a glycoprotein that specifically binds to gonocytes or Type A spermatogonia, on testis germ cell depletion were investigated by direct injection into the rete testis of a male camel. Seven recipients were prepared with DBA treatment, two males were castrated at 4 weeks for depletion assessment and the remaining five received donor cells 4-6 weeks after treatment. On average, similar to 17 million cells were isolated per gram of testis tissue, with 19.5 +/- 1.9% DBA-positive (DBA(+)) cells. Percoll centrifugation yielded a 1.5-fold increase in DBA(+) cells while Bovicoll centrifugation produced a 2.5-fold increase from the input cells of 18.6 +/- 2.1% DBA(+) cells. Semen was collected from the recipients 13-20 weeks after transfer and the presence of donor DNA in the samples was determined using microsatellite markers. In two of the five recipients, all semen samples were shown to be positive for donor-derived cells. These results demonstrate for the first time that: (1) heterologous testicular germ cell transplantation in camels is feasible and the recipients are able to produce spermatozoa of donor origin and (2) DBA can be used effectively to deplete endogenous stem cells.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据