title: Testicular injection of busulfan for recipient preparation in transplantation of spermatogonial stem cells in mice.
abstract: Intraperitoneal busulfan injections are used to prepare recipients for spermatogonial stem cell (SSC) transplantation but they are associated with haematopoietic toxicity. Testicular injections of busulfan have been proposed to overcome this limitation. To date, testicular injections have not been studied in the mouse model. Therefore, in the present study we used ICR mice as recipients for SSC transplantation and prepared these mice by testicular injection of busulfan on both sides (2, 3, 4 or 6mgkg-1per side). Following this, donor germ cells expressing red fluorescent protein (RFP) from transgenic C57BL/6J male mice were transplanted into recipients via the efferent duct on Days 16-17 after busulfan treatment. Positive control mice were prepared by intraperitoneal injection of 40mgkg-1busulfan and negative control mice were treated with bilateral testicular injection of 50% dimethyl sulfoxide. On Day 49 after transplantation, recipient mice that were RFP-positive by in vivo imaging were mated with ICR female mice. Donor-derived germ cell colonies with red fluorescence were observed on Day 60 after transplantation, and donor-derived offspring were obtained. The results demonstrated that endogenous germ cells were successfully eliminated in the seminiferous tubules via testicular busulfan administration, and that exogenous SSCs successfully undergo spermatogenesis in the testes of recipient mice prepared by testicular injections of busulfan. In addition to its effects on recipient preparation, this method was safe in rodents and could possibly be adapted for use in other species.
authors.affiliation: The Key Laboratory for Farm Animal Genetic Resources and Utilisation of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China.
authors.forename: Yusheng
authors.initials: Y
authors.lastname: Qin
authors.affiliation: The Key Laboratory for Farm Animal Genetic Resources and Utilisation of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Liu
authors.affiliation: The Key Laboratory for Farm Animal Genetic Resources and Utilisation of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China.
authors.forename: Yanan
authors.initials: Y
authors.lastname: He
authors.affiliation: Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education and Key Laboratory of Reproduction and Heredity of Ningxia Hui Autonomous Region, Ningxia Medical University, Yinchuan, 750004, China.
authors.forename: Wenzhi
authors.initials: W
authors.lastname: Ma
authors.affiliation: The Key Laboratory for Farm Animal Genetic Resources and Utilisation of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China.
authors.forename: Huabin
authors.initials: H
authors.lastname: Zhu
authors.affiliation: College of Animal Science and Technology, Jilin Agriculture University, Changchun, 130118, China.
authors.forename: Mingyuan
authors.initials: M
authors.lastname: Liang
authors.affiliation: The Key Laboratory for Farm Animal Genetic Resources and Utilisation of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China.
authors.forename: Haisheng
authors.initials: H
authors.lastname: Hao
authors.affiliation: The Key Laboratory for Farm Animal Genetic Resources and Utilisation of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China.
authors.forename: Tong
authors.initials: T
authors.lastname: Qin
authors.affiliation: The Key Laboratory for Farm Animal Genetic Resources and Utilisation of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China.
authors.forename: Xueming
authors.initials: X
authors.lastname: Zhao
authors.affiliation: The Key Laboratory for Farm Animal Genetic Resources and Utilisation of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China.
authors.forename: Dong
authors.initials: D
authors.lastname: Wang
mesh_terms: D000818:Animals; D002066:Busulfan / Q000008:administration & dosage*; D005260:Female; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D013091:Spermatogenesis; D013093:Spermatogonia / Q000637:transplantation*; D033581:Stem Cell Transplantation; D013737:Testis / Q000187:drug effects*
chemical_list: D002066:Busulfan