Derivation and propagation of spermatogonial stem cells from human pluripotent cells

Stem Cell Research & Therapy

Chencheng Yao¹  Zheng Li¹  Erlei Zhi¹  Peng Li¹  Ruhui Tian¹  Zijue Zhu¹  Chenhao Zhou¹  Shi Yang²  Minghui Niu³  Mengbo Yang³  Qingqing Yuan³  Zuping He³  Linhong Liu³  Yonghui Wang³  Wei-Qiang Gao⁴  Huiming Xu⁴ 

[1] Department of Andrology, the Center for Men’s Health, Urologic Medical Center, Shanghai Key Laboratory of Reproductive Medicine, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 100 Haining Road, 200080, Shanghai, China;Department of Urology, Shanghai Human Sperm Bank, Shanghai Institute of Andrology, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, 200135, Shanghai, China;State Key Laboratory of Oncogenes and Related Genes, Renji-MedX Clinical Stem Cell Research Center, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, 160 Pujian Road, 200127, Shanghai, China;State Key Laboratory of Oncogenes and Related Genes, Renji-MedX Clinical Stem Cell Research Center, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, 160 Pujian Road, 200127, Shanghai, China;School of Biomedical Engineering & Med-X Research Institute, Shanghai Jiao Tong University, Shanghai, China;

关键词: Human pluripotent stem cells;    Differentiation;    Human spermatogonia stem cells;    Xeno-transplantation;   

DOI  :  10.1186/s13287-020-01896-0

来源: Springer

【 摘 要 】

ObjectivesThis study is designed to generate and propagate human spermatogonial stem cells (SSCs) derived from human pluripotent stem cells (hPSCs).MethodshPSCs were differentiated into SSC-like cells (SSCLCs) by a three-step strategy. The biological characteristics of SSCLCs were detected by immunostaining with antibodies against SSC markers. The ability of self-renewal was measured by propagating for a long time and still maintaining SSCs morphological property. The differentiation potential of SSCLCs was determined by the generation of spermatocytes and haploid cells, which were identified by immunostaining and flow cytometry. The transcriptome analysis of SSCLCs was performed by RNA sequencing. The biological function of SSCLCs was assessed by xeno-transplantation into busulfan-treated mouse testes.ResultsSSCLCs were efficiently generated by a 3-step strategy. The SSCLCs displayed a grape-like morphology and expressed SSC markers. Moreover, SSCLCs could be propagated for approximately 4 months and still maintained their morphological properties. Furthermore, SSCLCs could differentiate into spermatocytes and haploid cells. In addition, SSCLCs displayed a similar gene expression pattern as human GPR125+ spermatogonia derived from human testicular tissues. And more, SSCLCs could survive and home at the base membrane of seminiferous tubules.ConclusionSSCLCs were successfully derived from hPSCs and propagated for a long time. The SSCLCs resembled their counterpart human GPR125+ spermatogonia, as evidenced by the grape-like morphology, transcriptome, homing, and functional characteristics. Therefore, hPSC-derived SSCLCs may provide a reliable cell source for studying human SSCs biological properties, disease modeling, and drug toxicity screening.

【 授权许可】

CC BY