ORIGINAL RESEARCH article
Front. Cell Dev. Biol.
Sec. Molecular and Cellular Reproduction
Volume 13 - 2025 | doi: 10.3389/fcell.2025.1634347
Integrating Microarray Data and Single-Cell RNA-Seq Reveals Correlation between Kit and Nmyc in Mouse Spermatogonia Stem Cell Population
Provisionally accepted
- 1University of Tehran, Tehran, Iran
- 2Amol University of Special Modern Technologies, Amol, Iran
- 3Universitat Heidelberg, Heidelberg, Germany
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Spermatogonial stem cells (SSCs) are essential for the continuous production of sperm and the maintenance of male fertility. Their selection, culture, and molecular characterization provide critical insights into spermatogenesis and potential therapeutic applications for male infertility. This study utilized CD49f-MACS and matrix selection techniques to isolate SSCs from mouse testicular samples. The molecular profile of the selected SSCs was analyzed through immunocytochemistry, gene ontology enrichment, weighted gene co-expression network analysis (WGCNA), and single-cell RNA sequencing (scRNA-seq). Additionally, protein-protein interaction (PPI) networks were constructed to identify key regulatory factors in SSC maintenance and differentiation. The selected SSCs exhibited a distinct molecular signature, with high expression of Dazl, Pou5f1 (Oct4), Gfra1, Nanog, and Kit. The Kit gene (c-kit) emerged as a crucial regulator of SSC differentiation, strongly associated with retinoic acid (RA)-mediated signaling pathways. Co-expression analysis revealed significant interactions between Kit, Nmyc, and other pluripotency-associated genes, highlighting its role in SSC development. Furthermore, single-cell RNA sequencing confirmed the dynamic expression of Kit during SSC differentiation and early meiosis initiation. Our findings underscore the pivotal role of Kit in spermatogenesis, reinforcing its potential as a therapeutic target for treating male infertility. The study also provides a comprehensive molecular framework for understanding SSC biology, with implications for regenerative medicine, fertility preservation, and in vitro gametogenesis. Further research integrating gene-editing technologies and in vivo models will be essential to explore the full therapeutic potential of SSC-based treatments.
Keywords: spermatogonial stem cells, KIT gene, Spermatogenesis, stem cell differentiation, Microarray, Pluripotency pathways
Received: 24 May 2025; Accepted: 18 Aug 2025.
Copyright: © 2025 Hashemi Karoii, Azizi and Skutella. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Hossein Azizi, Amol University of Special Modern Technologies, Amol, Iran
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