Differentiation of single cell derived human mesenchymal stem cells into cells with a neuronal phenotype: RNA and microRNA expression profile

Abstract

The adult bone marrow contains a subset of non-haematopoietic cells referred to as bone marrow mesenchymal stem cells (BMSCs). Mesenchymal stem cells (MSCs) have attracted immense research interest in the field of regenerative medicine due to their ability to be cultured for successive passages and multi-lineage differentiation. The molecular mechanisms governing the self-renewal and differentiation of MSCs remain largely unknown. In a previous paper we demonstrated the ability to induce human clonal MSCs to differentiate into cells with a neuronal phenotype (DMSCs). In the present study we evaluated gene expression profiles by Sequential Analysis of Gene Expression (SAGE) and microRNA expression profiles before and after the neuronal differentiation process. Various tissue-specific genes were weakly expressed in MSCs, including those of non-mesodermal origin, suggesting multiple potential tissue-specific differentiation, as well as stemness markers. Expression of OCT4, KLF4 and c-Myc cell reprogramming factors, which are modulated during the differentiation process, was also observed. Many peculiar nervous tissue genes were expressed at a high level in DMSCs, along with genes related to apoptosis. MicroRNA profiling and correlation with mRNA expression profiles allowed us to identify putative important genes and microRNAs involved in the differentiation of MSCs into neuronal-like cells. The profound difference in gene and microRNA expression patterns between MSCs and DMSCs indicates a real functional change during differentiation from MSCs to DMSCs.

G-CSF Receptor Positive Neuroblastoma Subpopulations Are Enriched in Chemotherapy-Resistant or Relapsed Tumors and Are Highly Tumorigenic

“…5B). Additional studies demonstrate that number of CD114+ upregulated microRNAs (miR-25, 106b, 17, 18, 19, 20a, 143, 27) are repressed upon neuronal lineage differentiation of somatic stem cells (33, 35, 36). In addition, microRNA pathway analysis with two independent prediction platforms (DIANA (DIANA Lab, Athens, Greece) and Ingenuity (Ingenuity Systems, Redwood City, CA) each demonstrated enrichment of p53, NGF, Notch and additional signaling pathways driving differentiation of neuronal and neural crest lineages (Supplementary Table S4 and Supplementary Table S5).…”

“…MicroRNAs play an important role in reprogramming and stem cell maintenance and recent studies have identified a number of microRNAs which regulate neuronal lineage specification from embryonic precursors (33, 34). To evaluate differential microRNA expression, we performed small-RNA sequencing on RNA directly isolated from FACS-purified CD114+ and CD114− subpopulations from three neuroblastoma cell lines (NGP, IMR-32, LA-N-5).…”

“…This is consistent with the resistance to apoptosis upon withdrawal of trophic factors observed in neuroblastoma precursors isolated from sympathetic ganglia of TH-MYCN transgenic mice (14). The CD114+ subpopulation displays increased expression of multiple microRNAs with validated targets in the p53 apoptotic response pathway, as well as microRNAs which promote reprogramming (44), stem cell cell-cycle regulation (30), and neuronal differentiation (33). Interestingly, repression of p53 through STAT3 activation appears to contribute to tumorigenesis (45), and small molecules which activate p53 and repress STAT3 can have potent antitumor effects (46).…”

Medical therapies with adult stem/progenitor cells (MSCs): A backward journey from dramatic results in vivo to the cellular and molecular explanations

“…Recently, there have been multiple reports that MSCs engage in extensive exchange of vesicles containing micro-RNAs during differentiation of the cells (Crobu et al, 2011; Zhang et al, 2011) and in their interaction with malignant cells (Gregory et al, 2011). (For recent review see Guo et al, 2011).…”

Extracellular matrix-regulated neural differentiation of human multipotent marrow progenitor cells enhances functional recovery after spinal cord injury

“…Tenascin-cytotactin (TN-C), highly expressed in the subventricular zone, generates a stem cell “niche” that acts to orchestrate neural stem cell development in mice [23,24]. Among transdifferentiation studies, most have been conducted with animal cells [25–28] and some with human MPCs [29,30]. …”