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Keywords
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Spermatogonial stem cells, Embryonic stem-like cells, Microarray analysis, PPI network, Signaling
pathways, miRNA and lncRNA
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Abstract
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With the significant challenges in using human embryonic stem cells (ESCs) for research and clinical applications,
there is a growing impetus to seek alternative pluripotent cell sources. Embryonic stem-like (ES-like) cells emerge
as a promising avenue in this pursuit. Our research demonstrates the potential for deriving ES-like cells from spermatogonial
stem cells (SSCs) in a time-dependent manner under defined culture conditions. To better understand
this process, we investigated the gene expression dynamics and underlying pathways associated with ES-like cell
generation from SSCs. A deeper understanding of the signaling pathways underlying this biological process can lead
us to refine protocols for ES-like cell generation, which could catalyze the development of more efficient and expedited
methodologies inspired by the derivation pathway for future research in regenerative medicine. To identify
differentially expressed genes (DEGs), we analyzed publicly available microarray data from murine cells obtained
from the Gene Expression Omnibus (GEO). This analysis enabled the prediction of protein–protein interactions (PPIs),
which were subsequently used for pathway enrichment analysis to identify biologically relevant pathways. Complementing
these computational findings, we conducted in vitro experiments, including Fluidigm qPCR and immunostaining.
These experiments serve as validation for our microarray data and the DEGs identified, providing reassurance
about the reliability of our research. Among the identified enriched pathways in our investigation are the Toll-like
receptor (TLR), GDNF/RET, interleukins (ILs), FGF/FGFR, and SMAD signaling pathway, along with the activation
of NIMA kinases. Additionally, miR-410-3p, miRNA let-7e, Miat, and Xist are among some of the predicted non-coding
RNAs.
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