期刊
JOURNAL OF CELLULAR AND MOLECULAR MEDICINE
卷 26, 期 5, 页码 1567-1578出版社
WILEY
DOI: 10.1111/jcmm.17197
关键词
germ cell transplantation; in vitro culture; Rhesus monkey; spermatogonia; spermatogonial stem cells
资金
- National Key Research and Development Program of China [2018YFA0801403, 2018YFC1003302]
- National Natural Science Foundation of China [2021YFC2700201, 32070831, 31872844]
This study characterized germ cells in macaque testes and successfully isolated spermatogenic cells with high purity. Autologous cell transplantation in macaque testes was mimicked under ultrasonographic guidance. Although SSC propagation in vitro was not significant, monkey testicular germ cells with stem cell characteristics were maintained.
Fertility preservation for prepubertal cancer patients prior to oncologic treatment is an emerging issue, and non-human primates are considered to constitute suitable models due to the limited availability of human testicular tissues. However, the feasibility of spermatogonial stem cell (SSC) propagation in vitro and autologous testicular germ cell transplantation in vivo requires further exploration in monkeys. Herein, we characterized germ cells in macaque testes at 6 months (M), 18 M and 60 M of age, and effectively isolated the spermatogenic cells (including the spermatogonia) from macaque testes with high purity (over 80%) using combined approaches of STA-PUT separation, Percoll gradients and differential plating. We also generated recipient monkey testes with ablated endogenous spermatogenesis using the alkylating agent busulfan in six macaques, and successfully mimicked autologous cell transplantation in the testes under ultrasonographic guidance. The use of trypan blue led to successful intratubular injection in 4 of 4 testes. Although SSCs in culture showed no significant propagation, we were able to maintain monkey testicular germ cells with stem cell characteristics for up to 3 weeks. Collectively, these data provided meaningful information for future fertility preservation and SSC studies on both non-human primates and humans.
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