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

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 transcriptomic 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 transcription factors were upregulated, and 45 were downregulated in NPCs. Neurogenesis related genes were upregulated 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 enrichment analysis by ChEA3 revealed Foxs1 and HEYL, amongst the top five transcription factors, inhibits and enhances, respectively, the NPCs differentiation of MSCs. Conclusions The vast differences in the transcriptomic profiles between NPCs and MSCs revealed a set of markers that can identify the differentiation stage of NPCs as well as provide new targets to enhance MSCs differentiation into NPCs.

Challenges and advances in clinical applications of mesenchymal stromal cells

“…Moreover, MSCs have shown the capacity to differentiate into numerous cell types such as neural cells, hepatocyte-like cells, and pancreatic islet-like cells [89,90]. The transient differentiation of MSCs into neural precursor-like cells may experience de-differentiation during extended culture [91].…”

Biomaterials as a Vital Frontier for Stem Cell-Based Tissue Regeneration

“…Furthermore, MSC can differentiate into cells derived from other embryonic lineages, based on the culture conditions, with variable degrees of success. For example, MSC was shown to differentiate into several types of cells including hepatocytes, beta cell of the pancreas, neurons and neuroectodermal cells ( Cooke M. J. et al, 2010 ; Elsharkawi et al, 2019 ; Mansour et al, 2019 ; Khan et al, 2020 ). In-vitro , the proliferation of ASC, is limited by Hayflick’s limit, associated with senescence and decreasing ability to differentiate following few passages in culture.…”

Chitosan Hydrogel Supplemented with Metformin Promotes Neuron–like Cell Differentiation of Gingival Mesenchymal Stem Cells

“…The number of new neurons in mice with neurological damage increased after taking metformin [ 14 ]. Growth factors, such as (Epidermal Growth Factor) EGF, bFGF [ 15 ], and brain–derived neurotrophic factor (BDNF) [ 16 ], can effect cell growth, development, and differentiation, and they can regulate pathways by activating the receptors on the cell surface. Growth factors are often expressed in nerve tissues, can induce differentiation of neural stem cells and promote proliferation and development, and have neuroprotective effects.…”