Profiling of MicroRNA in Human and Mouse ES and iPS Cells Reveals Overlapping but Distinct MicroRNA Expression Patterns

Abstract

Using quantitative PCR-based miRNA arrays, we comprehensively analyzed the expression profiles of miRNAs in human and mouse embryonic stem (ES), induced pluripotent stem (iPS), and somatic cells. Immature pluripotent cells were purified using SSEA-1 or SSEA-4 and were used for miRNA profiling. Hierarchical clustering and consensus clustering by nonnegative matrix factorization showed two major clusters, human ES/iPS cells and other cell groups, as previously reported. Principal components analysis (PCA) to identify miRNAs that segregate in these two groups identified miR-187, 299-3p, 499-5p, 628-5p, and 888 as new miRNAs that specifically characterize human ES/iPS cells. Detailed direct comparisons of miRNA expression levels in human ES and iPS cells showed that several miRNAs included in the chromosome 19 miRNA cluster were more strongly expressed in iPS cells than in ES cells. Similar analysis was conducted with mouse ES/iPS cells and somatic cells, and several miRNAs that had not been reported to be expressed in mouse ES/iPS cells were suggested to be ES/iPS cell-specific miRNAs by PCA. Comparison of the average expression levels of miRNAs in ES/iPS cells in humans and mice showed quite similar expression patterns of human/mouse miRNAs. However, several mouse- or human-specific miRNAs are ranked as high expressers. Time course tracing of miRNA levels during embryoid body formation revealed drastic and different patterns of changes in their levels. In summary, our miRNA expression profiling encompassing human and mouse ES and iPS cells gave various perspectives in understanding the miRNA core regulatory networks regulating pluripotent cells characteristics.

Identification of RNA-Binding Protein LARP4B as a Tumor Suppressor in Glioma

“…We used two NSC lines differentiated from two human iPS cells (tkCB_Sev9 and tkDA_3-4; ref. 19) and one commercially available NSC line derived from an embryonic stem cell (ES) line. We also used three glioma stem cells (14) and six glioma cell lines.…”

Changes in microRNA expression during differentiation of embryonic and induced pluripotent stem cells to definitive endoderm

“…PCR reactions were 21 carried out using the following PCR cycling conditions: 10min at 95°C; followed by 40 22 repeated cycles of 5sec at 95°C, 15sec at 58°C and 10sec at 72°C. The fluorescence was 23 acquired at the end of the elongation step. mRNA expression levels were normalised against 24 the CYCG reference gene.…”

“…This suggests that miRNAs which are likely to be co-regulated and co-transcribed 20 show similar expression patterns in these samples, indicating that the results of the 21 microarray follow expected biological patterns, offering confidence in the validity of the 22 results. 23 To aid in the identification of candidate miRNAs that may be important in the differentiation 24 of iPSCs and ESCs into DE, we decided to analyse the miRNA expression patterns in four 25 distinct sample populations as they differentiated into DE: iPSCs (MRC5I and MRC9G 26 combined), ESCs (H1 and H9 combined) and, based on the fact that the hierarchical 27 clustering analysis identified the H1 and H9 ESC lines as clearly distinct populations, we also 28 assessed these ESC lines individually. 86 miRNAs were identified as being differentially 29 expressed between differentiated and undifferentiated iPSCs, while 82 miRNAs were 30 differentially expressed between differentiated and undifferentiated ESCs.…”

“…Instead, most of these studies describe the generation of cells 23 that were functionally restricted, either having a polyhormonal phenotype, low levels of 24 insulin synthesis, or lacking appropriate insulin release in response to high glucose. It has 25 been suggested that these characteristics are more reminiscent of foetal pancreatic cells (2) 26 and that a more complete maturation into a pancreatic β-cell phenotype is only possible after 27 the transplantation of pancreatic progenitor cells into immune-deficient mice, where as yet 28 unknown factors in the in vivo environment are capable of inducing these cells to more 29 complete differentiation (3; 4; 5).…”

<b>Estudo do potencial de pluripotência de células-tronco equinas derivadas de tecido adulto e cordão umbilical submetidas à reprogramação induzida geneticamente (células iPS)</b>

“…Têm importante papel na manutenção de uma célula, controlando funções essenciais como proliferação e morte, podendo ainda atuar como oncogenes, como supressores de tumor ou ainda como ambos DUTTA, 2009;ROMANO et al, 2012;SANDMAIER;TELUGO, 2015). O perfil de miRNAs de células iPS já foi analisado em outras espécies como humanos (WILSON et al, 2009;RAZAK et al, 2013;GREENE et al, 2014;WANG et al, 2015), murinos (PORCIUNCULA et al, 2013) e suínos .…”