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Adaptation of Human Testicular Niche Cells for Pluripotent Stem Cell and Testis Development Research

  • Original Article
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Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Background:

Human testicular cells are greatly valuable to the research community as tools for studying testicular physiology and the effects of environmental pollutants. Because adult testicular cells have a limited self-organization capacity and life span, we investigated whether human pluripotent stem cells (hPSCs) can be used together with testicular cells to move a step closer toward making an optimal model of the human testis.

Methods:

We used in vitro culture of donor testicular cells under serum-containing and chemically defined conditions. CRISPR-Cas9 technology was applied to introduce fluorescent transgenes (mCherry2 and EGFP) into hPSCs and testicular cells. hPSC-derived spheroids were co-cultured with human testicular cells in mini-spin bioreactors.

Results:

Traditional cell culture conditions used for maintenance of testicular somatic cells generally contain serum and pose limitations on evaluating the role of active molecules on cell functions. We established that chemically defined culture conditions can be used to maintain testicular cells without the loss of proliferative activity. These cultures demonstrate marker expression which is characteristic of common testicular cell types: Sertoli, Leydig, endothelial, myoid cells, and macrophages. In order to model testicular physiology, it is important to be able to perform live cell microscopy. Thus, we generated fluorescent protein-expressing human testicular cells and hPSCs and demonstrated that these cell types can be successfully co-cultured for prolonged periods of time in a three-dimensional microenvironment.

Conclusion:

Our research extends the possible applications of human testis-derived somatic cells and shows that they can be used together with hPSCs for further studies of human male reproductive biology.

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Acknowledgements

We thank Immanuel Rasool from WRTC for coordinating the acquisition of deidentified human testis samples used for this study, Stephen Wellard for technical assistance, Ran Brosh for providing the EF1α promoter sequence, and Barry Zirkin and JinYong Chung for discussion. This work was supported by a KY Cha Award In Stem Cell Technology from the American Society for Reproductive Medicine and a research grant from National Institute of General Medical Sciences (R01GM11755), both awarded to P.W.J.

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  1. Department of Biochemistry and Molecular Biology, Johns Hopkins University Bloomberg School of Public Health, 615 N. Wolfe St., Baltimore, MD, 21205, USA

    Marina V. Pryzhkova & Philip W. Jordan

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  1. Marina V. Pryzhkova
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Correspondence to Philip W. Jordan.

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Ethical statement

The use of human ESC line was approved by the JHU institutional stem cell research oversight committee (protocol ISCRO00000089). Deidentified donor testes tissues were designated as “not human subjects research” by Johns Hopkins University (JHU IRB No: 00006700).

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Pryzhkova, M.V., Jordan, P.W. Adaptation of Human Testicular Niche Cells for Pluripotent Stem Cell and Testis Development Research. Tissue Eng Regen Med 17, 223–235 (2020). https://doi.org/10.1007/s13770-020-00240-0

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