Significant transcriptomic changes are associated with differentiation of bone marrow-derived mesenchymal stem cells int
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Cell & Bioscience Open Access
RESEARCH
Significant transcriptomic changes are associated with differentiation of bone marrow‑derived mesenchymal stem cells into neural progenitor‑like cells in the presence of bFGF and EGF Amir Ali Khan1,5*† , Tee Jong Huat2,3†, Abdullah Al Mutery1, Ahmed Taher El‑Serafi4, Hassen Hadj Kacem1,5, Sallam Hasan Abdallah5, Muhammed Faruque Reza3, Jafri Malin Abdullah3,6 and Hasnan Jaafar7
Abstract Introduction: Mesenchymal stem cells (MSCs) isolated from bone marrow have different developmental origins, including neural crest. MSCs can differentiate into neural progenitor-like cells (NPCs) under the influence of bFGF and EGF. NPCs can terminally differentiate into neurons that express beta-III-tubulin and elicit action potential. The main aim of the study was to identify key genetic markers involved in differentiation of MSCs into NPCs through transcrip‑ tomic analysis. Method: Total RNA was isolated from MSCs and MSCs-derived NPCs followed by cDNA library construction for transcriptomic analysis. Sample libraries that passed the quality and quantity assessments were subjected to high throughput mRNA sequencing using NextSeq®500. Differential gene expression analysis was performed using the DESeq2 R package with MSC samples being a reference group. The expression of eight differentially regulated genes was counter validated using real-time PCR. Results: In total, of the 3,252 differentially regulated genes between MSCs and NPCs with two or more folds, 1,771 were upregulated genes, whereas 1,481 were downregulated in NPCs. Amongst these differential genes, 104 tran‑ scription factors were upregulated, and 45 were downregulated in NPCs. Neurogenesis related genes were upregu‑ lated in NPCs and the main non-redundant gene ontology (GO) terms enriched in NPCs were the autonomic nervous system, cell surface receptor signalling pathways), extracellular structure organisation, and programmed cell death. The main non-redundant GO terms enriched in MSCs included cytoskeleton organisation cytoskeleton structural constituent, mitotic cell cycle), and the mitotic cell cycle process Gene set enrichment analysis also confirmed cell cycle regulated pathways as well as Biocarta integrin pathway were upregulated in MSCs. Transcription factors enrich‑ ment analysis by ChEA3 revealed Foxs1 and HEYL, amongst the top five transcription factors, inhibits and enhances, respectively, the NPCs differentiation of MSCs.
*Correspondence: [email protected] † Amir Ali Khan and Tee Jong Huat contributed equally to this work 1 Department of Applied Biology, College of Sciences, University of Sharjah, P.O. Box 27272, Emirate of Sharjah, United Arab Emirates Full list of author information is available at the end of the article © The Author(s) 2020. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, p
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