MicroRNA signatures of iPSCs and endoderm-derived tissues

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

“…Washing was carried out according to the 11 manufacturer's instructions. On-column DNase digestion was performed using the RNase- 12 free DNase kit (Qiagen). Total RNA (including the miRNA fraction) was eluted in 30µl 13 water.…”

“…48h after plasmid transfection, the supernatant was collected and analysed for 15 luminescence using the SecretePair™ Dual Luminescence Assay Kit (Genecopoeia) 16 according to the manufacturer's instructions. 21 Prior to differentiation experiments, we derived two iPSC lines using methods previously using the two iPSC lines and the H9 cell line, which is a well-characterised ESC line, we 12 recognised that since H9 was grown on Matrigel™ and the iPSCs were grown on iSNL 13 feeders, the inclusion of an ESC line that was grown on feeders may reveal additional 14 miRNAs of interest. For this reason H1 was introduced into the microarray analysis.…”

“…As a result, miRNAs are likely to have a considerable 11 impact on the rapid and efficient control of gene expression. 12 Conditional deletion of the miRNA processing enzyme DICER-1 in the developing pancreas 13 results in defects in all pancreatic lineages, but it particularly affects the insulin-secreting β-14 cells (8), demonstrating the critical role that miRNAs play in pancreatic development. While 15 there have been a number of reports describing the miRNA expression at the later stages of 16 pancreatic development (reviewed in Francis et al) (9), only a small number of studies have 17 investigated miRNA expression at the definitive endoderm (DE) stage (10; 11; 12) which has 18 previously been identified as critical in the in vitro differentiation towards a pancreatic 19 lineage (13).…”

<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 .…”

“…A família de miRNA let-7 é conhecida por ser uma das primeiras famílias miRNAs identificadas e mais intensamente estudados em humanos, é um grupo de miRNAs pró-diferenciação e alterações na sua expressão estão associadas a canceres agressivos (ROUSH;SLACK, 2008;SU et al, 2012;PORCIUNCULA et al, 2013 É importante ressaltar que os miRNAs não só são relevantes para a regulação da reprogramação e manutenção da pluripotência, como podem apresentar padrões de expressão variados, de acordo com diferentes estados de pluripotência .…”