Primary culture of germ cells that portray stem cell characteristics and recipient preparation for autologous transplantation in the rhesus monkey

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) propag...

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Bibliographic Details
Published in:Journal of cellular and molecular medicine 2022-03, Vol.26 (5), p.1567-1578
Main Authors: Yuan, Huaqin, Sun, Jiachen, Wang, Shengnan, Xiang, Ziyi, Yang, Fan, Yan, Yaping, Duan, Yanchao, Li, Lufan, Wu, Xin, Si, Wei
Format: Article
Language:English
Subjects:
Animal models
Animals
Antibodies
Autografts
Biopsy
Busulfan
Cell culture
Cryopreservation
Enzymes
Fertility
Gene expression
germ cell transplantation
Germ cells
Histology
Humans
in vitro culture
Macaca mulatta
Male
Microscopy
Monkeys & apes
Original
Preservation
Propagation
Rhesus monkey
Sperm
Spermatogenesis
Spermatogonia
spermatogonial stem cells
Stem Cell Transplantation
Stem Cells
Testes
Testis
Transplantation
Transplantation, Autologous
Ultrasonic imaging
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Summary: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.
ISSN:1582-1838
1582-4934
DOI:10.1111/jcmm.17197