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Article

ITGA6+ Human Testicular Cell Populations Acquire a Mesenchymal Rather than Germ Cell Transcriptional Signature during Long-Term Culture

1
Reproductive Biology Laboratory, Center for Reproductive Medicine, Amsterdam UMC, Amsterdam Reproduction & Development Research Institute, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands
2
Department of Epidemiology & Data Science, Amsterdam UMC, Amsterdam Public Health Research Institute, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands
3
Amsterdam UMC, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(21), 8269; https://doi.org/10.3390/ijms21218269
Received: 25 September 2020 / Revised: 26 October 2020 / Accepted: 28 October 2020 / Published: 4 November 2020
(This article belongs to the Special Issue Molecular Basis of Fertility Preservation and Restoration 2.0)
Autologous spermatogonial stem cell transplantation is an experimental technique aimed at restoring fertility in infertile men. Although effective in animal models, in vitro propagation of human spermatogonia prior to transplantation has proven to be difficult. A major limiting factor is endogenous somatic testicular cell overgrowth during long-term culture. This makes the culture both inefficient and necessitates highly specific cell sorting strategies in order to enrich cultured germ cell fractions prior to transplantation. Here, we employed RNA-Seq to determine cell type composition in sorted integrin alpha-6 (ITGA6+) primary human testicular cells (n = 4 donors) cultured for up to two months, using differential gene expression and cell deconvolution analyses. Our data and analyses reveal that long-term cultured ITGA6+ testicular cells are composed mainly of cells expressing markers of peritubular myoid cells, (progenitor) Leydig cells, fibroblasts and mesenchymal stromal cells and only a limited percentage of spermatogonial cells as compared to their uncultured counterparts. These findings provide valuable insights into the cell type composition of cultured human ITGA6+ testicular cells during in vitro propagation and may serve as a basis for optimizing future cell sorting strategies as well as optimizing the current human testicular cell culture system for clinical use. View Full-Text
Keywords: fertility preservation; spermatogonia; stem cells; bulk transcriptome; germ cells; cell culture; cell type deconvolution fertility preservation; spermatogonia; stem cells; bulk transcriptome; germ cells; cell culture; cell type deconvolution
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MDPI and ACS Style

Struijk, R.B.; Mulder, C.L.; van Daalen, S.K.M.; de Winter-Korver, C.M.; Jongejan, A.; Repping, S.; van Pelt, A.M.M. ITGA6+ Human Testicular Cell Populations Acquire a Mesenchymal Rather than Germ Cell Transcriptional Signature during Long-Term Culture. Int. J. Mol. Sci. 2020, 21, 8269. https://doi.org/10.3390/ijms21218269

AMA Style

Struijk RB, Mulder CL, van Daalen SKM, de Winter-Korver CM, Jongejan A, Repping S, van Pelt AMM. ITGA6+ Human Testicular Cell Populations Acquire a Mesenchymal Rather than Germ Cell Transcriptional Signature during Long-Term Culture. International Journal of Molecular Sciences. 2020; 21(21):8269. https://doi.org/10.3390/ijms21218269

Chicago/Turabian Style

Struijk, Robert B., Callista L. Mulder, Saskia K.M. van Daalen, Cindy M. de Winter-Korver, Aldo Jongejan, Sjoerd Repping, and Ans M.M. van Pelt. 2020. "ITGA6+ Human Testicular Cell Populations Acquire a Mesenchymal Rather than Germ Cell Transcriptional Signature during Long-Term Culture" International Journal of Molecular Sciences 21, no. 21: 8269. https://doi.org/10.3390/ijms21218269

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