Abstract
MicroRNAs are key modulators at molecular level in different biological processes, including determination of cell fate and differentiation. Herein, microRNA expression profiling experiments were performed on syngeneic cardiac (CStC) and bone marrow (BMStC) mesenchymal stromal cells cultured in standard growth medium and then in vitro exposed to adipogenic, osteogenic, cardiomyogenic and endothelial differentiation media. Analysis identified a tissue-specific microRNA signature composed of 16 microRNAs that univocally discriminated cell type of origin and that were completely unaffected by in vitro differentiation media: 4 microRNAs were over-expressed in cardiac stromal cells, and 12 were overexpressed or present only in bone marrow stromal cells. Further, results revealed microRNA subsets specifically modulated by each differentiation medium, irrespective of the cell type of origin, and a subset of 7 microRNAs that were down-regulated by all media with respect to growth medium. Finally, we identified 16 microRNAs that were differentially modulated by the media when comparing the two tissues of origin. The existence of a tissue-specific microRNA signature surviving to any differentiation stimuli, strongly support the role if microRNAs determining cell identity related to tissue origin. Moreover, we identified microRNA subsets modulated by different culture conditions in a tissue-specific manner, pointing out their importance during differentiation processes.
Highlights
MicroRNAs are 21–23 nucleotide non-coding RNA molecules, which modulate the stability and/or the translational efficiency of messenger RNAs
cardiac mesenchymal stromal cells (CStC) were enzymatically isolated from small auricle fragments using 3 mg/ml collagenase (Serva) and cultured in standard growth medium (GM), composed of Iscove’s Modified Dulbecco’s Medium (IMDM, Lonza) supplemented with 20% fetal bovine serum (FBS, Hyclone), 10 ng/ml basic Fibroblasts Growth Factor, 10.000 U/ml Penicillin/Streptomycin (Invitrogen), 20 mM L-Glutamine (Sigma-Aldrich)
We confirmed that CStC [30] displayed greater propensity to differentiate into cardiomyocyte-like and endothelial-like cells when compared to BMStC [31]
Summary
MicroRNAs (miRs) are 21–23 nucleotide non-coding RNA molecules, which modulate the stability and/or the translational efficiency of messenger RNAs (mRNA). MiR have been involved in pluripotency maintenance [4], cell proliferation and differentiation [5], epithelial to mesenchymal transition [6], senescence [7], and apoptosis [8]. Due to their wide role in cell process regulation, miR have gained popularity as tools that are able to promote direct cell to cell phenotypic conversion as well as adult cell reprogramming into pluripotent stem cells. Being able to regulate and, possibly, to fine tune cell fate, miRs appear as a new frontier for application in regenerative medicine
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