Meso-Endothelial bipotent progenitors from human placenta display distinct molecular and cellular identity
Then total RNA from CD45-CD34+CD31Neg, CD45-CD34+CD31Lo and CD45-CD34+CD31Int colonies at passage 0 was extracted using the RNA easy Plus Mini Kit (Qiagen, CA) according to manufacturer s instruction. The quality and quantity of isolated RNA was analysed with Agilent 2100 Bioanalyzer (Agilent Technologies, Mulgrave, Victoria). RNA-seq libraries were prepared using the manufacturer s instructions followed by sequencing on Illumina s HiSeq 2500 (Illumina, CA) performed at the Garvan Institute (Sydney, Australia). Sequence data were aligned with STAR (version 2.5.0c). The read counts were measured using the GENCODE gene annotation. Differential gene expression was measured with DESeq2. Genes with significant expression levels were selected by applied a false discovery rate-adjusted P value cut-off of P <0.05, and a log fold change cut-off of 2.