In Vitro Propagation of XXY Undifferentiated Mouse Spermatogonia: Model for Fertility Preservation in Klinefelter Syndrome Patients

Galdon, Guillermo; Deebel, Nicholas A.; Zarandi, Nima Pourhabibi; Pettenati, Mark J.; Kogan, Stanley; Wang, Christina; Swerdloff, Ronald S.; Atala, Anthony; Lue, Yanhe; Sadri-Ardekani, Hooman

doi:10.3390/ijms23010173

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In Vitro Propagation of XXY Undifferentiated Mouse Spermatogonia: Model for Fertility Preservation in Klinefelter Syndrome Patients

Guillermo Galdon

^1,2

Nicholas A. Deebel

^1,3,

Nima Pourhabibi Zarandi

¹,

⁴,

^1,3,

⁵,

⁵,

^1,3

⁵ and

Hooman Sadri-Ardekani

^1,3,*

Wake Forest Institute for Regenerative Medicine (WFIRM), Wake Forest School of Medicine, Winston-Salem, NC 27101, USA

Facultad de Medicina Universidad de Barcelona, 08036 Barcelona, Spain

Department of Urology, Wake Forest School of Medicine, Winston-Salem, NC 27101, USA

⁴

Section of Medical Genetics, Department of Pathology, Wake Forest School of Medicine, Winston-Salem, NC 27101, USA

⁵

Division of Endocrinology, The Lundquist Institute and Harbor-UCLA Medical Center, Torrance, CA 90502, USA

Author to whom correspondence should be addressed.

Academic Editor: Katja J. Teerds

Int. J. Mol. Sci. 2022, 23(1), 173; https://doi.org/10.3390/ijms23010173

Received: 16 October 2021 / Revised: 15 December 2021 / Accepted: 19 December 2021 / Published: 24 December 2021

(This article belongs to the Special Issue Molecular Basis of Fertility Preservation and Restoration)

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Abstract

Klinefelter syndrome (KS) is characterized by a masculine phenotype, supernumerary sex chromosomes (usually XXY), and spermatogonial stem cell (SSC) loss in their early life. Affecting 1 out of every 650 males born, KS is the most common genetic cause of male infertility, and new fertility preservation strategies are critically important for these patients. In this study, testes from 41, XXY prepubertal (3-day-old) mice were frozen-thawed. Isolated testicular cells were cultured and characterized by qPCR, digital PCR, and flow cytometry analyses. We demonstrated that SSCs survived and were able to be propagated with testicular somatic cells in culture for up to 120 days. DNA fluorescent in situ hybridization (FISH) showed the presence of XXY spermatogonia at the beginning of the culture and a variety of propagated XY, XX, and XXY spermatogonia at the end of the culture. These data provide the first evidence that an extra sex chromosome was lost during innate SSC culture, a crucial finding in treating KS patients for preserving and propagating SSCs for future sperm production, either in vitro or in vivo. This in vitro propagation system can be translated to clinical fertility preservation for KS patients. View Full-Text

Keywords: spermatogonia; spermatogonia stem cells; Klinefelter syndrome; male infertility; fertility preservation

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Galdon, G.; Deebel, N.A.; Zarandi, N.P.; Pettenati, M.J.; Kogan, S.; Wang, C.; Swerdloff, R.S.; Atala, A.; Lue, Y.; Sadri-Ardekani, H. In Vitro Propagation of XXY Undifferentiated Mouse Spermatogonia: Model for Fertility Preservation in Klinefelter Syndrome Patients. Int. J. Mol. Sci. 2022, 23, 173. https://doi.org/10.3390/ijms23010173

AMA Style

Galdon G, Deebel NA, Zarandi NP, Pettenati MJ, Kogan S, Wang C, Swerdloff RS, Atala A, Lue Y, Sadri-Ardekani H. In Vitro Propagation of XXY Undifferentiated Mouse Spermatogonia: Model for Fertility Preservation in Klinefelter Syndrome Patients. International Journal of Molecular Sciences. 2022; 23(1):173. https://doi.org/10.3390/ijms23010173

Chicago/Turabian Style

Galdon, Guillermo, Nicholas A. Deebel, Nima P. Zarandi, Mark J. Pettenati, Stanley Kogan, Christina Wang, Ronald S. Swerdloff, Anthony Atala, Yanhe Lue, and Hooman Sadri-Ardekani. 2022. "In Vitro Propagation of XXY Undifferentiated Mouse Spermatogonia: Model for Fertility Preservation in Klinefelter Syndrome Patients" International Journal of Molecular Sciences 23, no. 1: 173. https://doi.org/10.3390/ijms23010173

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