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Efficient Differentiation of Mouse Embryonic Stem Cells into Motor Neurons
Published on: June 9, 2012
Efficient differentiation of gonadal somatic cell-like cells (GSCLCs) using magnetic nanoparticles from mouse
Ullah Rahman1, Zhihui Liu1,2, Yuan Gao1
1Jiangsu Key Laboratory for Molecular and Medical Biotechnology, College of Life Sciences, Nanjing Normal University Nanjing 210046, Jiangsu, PR China.
Objectives:
Successful induction of gonadal somatic cell-like cells (GSCLCs) from mouse embryonic stem cells (mES) is important for studying ovarian somatic cell differentiation, development and their function for oocyte genesis and maturation. This study demonstrates the successful differentiation of GSCLCs from mES in vitro by utilizing novel green-synthesized Fe3O4 nanoparticles (G-Fe3O4-NP).
Methods:
mES were differentiated in vitro using G-Fe3O4-NP in the absence of BMP4, RA and SHH. GSCLCs identity was confirmed by RT-qPCR analysis of Foxl2, Amh, and Gata4, and by Immunofluorescence staining of FOXL2, AMH, FSHR, and GATA4. Mitochondrial function and cellular redox status were evaluated using JC-1 and DCFH-DA assay. Gene Set Enrichment analysis (GSEA) was performed to evaluate transcriptional reprogramming during GSCLCs differentiation.
Results:
G-Fe3O4-NP derived GSCLCs were achieved without supplementation of BMP4 and retinoic acid considered as key cellular factor for GSCLCs differentiation, suggesting a unique ability of G-Fe3O4-NP to independently induce gonadal somatic lineages. Transcriptional profiling and gene set enrichment analysis (GSEA) further elucidated the interplay of transcriptional, epigenetic, and metabolic reprogramming during GSCLC differentiation. Metabolic assessments showed that GSCLCs possessed functional mitochondria and maintain basal oxidative stress levels, reflecting mitochondrial functionality and stability. Key regulators including transcription factor 21 (TCF21), E-cadherin-mediated epithelial stabilization, and interleukin-12 were identified as a critical drivers of GSCLC maturation, supporting cells structural integrity, homeostasis, and gonadal lineage specification.
Conclusion:
This study establishes that G-Fe3O4-NPs enable the cytokine independent derivation of GSCLCs from mES by driving lineage specification. This coordinated transcriptional, epigenetic, and metabolic reprogramming establishes a foundation for in vitro GSCLCs generation.
