InVitro Meiosis of Male Germline Stem Cells.

Qijing Lei,Susana M Chuva De Sousa Lopes,Xin Lai,Ans M.M Van Pelt,Jitske Eliveld,Geert Hamer

doi:10.1016/j.stemcr.2020.10.006

Qijing Lei, Susana M Chuva De Sousa Lopes + Show 4 more

Open Access

https://doi.org/10.1016/j.stemcr.2020.10.006

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Abstract

SummaryIn vitro spermatogenesis has been achieved by culturing mouse embryonic stem cells (ESCs) together with a cell suspension of male juvenile gonad. However, for human fertility treatment or preservation, patient-specific ESCs or juvenile gonad is not available. We therefore aim to achieve in vitro spermatogenesis using male germline stem cells (GSCs) without the use of juvenile gonad. GSCs, when cultured on immortalized Sertoli cells, were able to enter meiosis, reach the meiotic metaphase stages, and sporadically form spermatid-like cells. However, the in vitro-formed pachytene-like spermatocytes did not display full chromosome synapsis and did not form meiotic crossovers. Despite this, the meiotic checkpoints that usually eliminate such cells to prevent genomic instabilities from being transmitted to the offspring were not activated, allowing the cells to proceed to the meiotic metaphase stages. In vitro-generated spermatid-like cells should thus be thoroughly investigated before being considered for clinical use.

Highlights

An estimated 10%–15% of couples suffer from subfertility, of which roughly 50% are diagnosed with male factor infertility (Kumar and Singh, 2015)
As published previously (Wang et al, 2016), we observed that, when induced to differentiate using RA and grown in vitro on a feeder layer of mouse embryonic fibroblasts (MEFs), germline stem cells (GSCs) can develop into zygotene spermatocytes and, occasionally, even form pachytene-like spermatocytes
During the first stage the GSCs were cultured with the factors known to be necessary for maintaining SSC self-renewal, glial-derived neurotrophic factor (GDNF), basic fibroblast growth factor, and epidermal growth factor (EGF) (Kanatsu-Shinohara et al, 2003), and behaved exactly as described previously (Zheng et al, 2018)

Summary

Introduction

An estimated 10%–15% of couples suffer from subfertility, of which roughly 50% are diagnosed with male factor infertility (Kumar and Singh, 2015). This was achieved by complete in vitro differentiation of embryonic stem cells (ESCs) (Easley IV et al, 2012; Geijsen et al, 2004; Nayernia et al, 2006; Zhou et al, 2016) or by first differentiating mouse ESCs to primordial germ cell-like cells (PGCLCs), spermatogonial stem celllike cells, or germline stem cell (GSC)-like cells in vitro, which can undergo spermatogenesis after transplantation into the seminiferous tubules of infertile mice (Hayashi et al, 2011; Ishikura et al, 2016; Li et al, 2019) All these studies have reported the generation of haploid spermatid-like cells, only one study (Zhou et al, 2016) was able to recapitulate most key events that characterize successful meiosis and generation of fertile haploid germ cells in vitro (Handel et al, 2014). Even in a future clinic, ESCs comprising the patient’s own genetic material will most likely not be available for an adult human patient

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