Single-Cell analysis of human iPS-derived Cells Undergoing Endothelial-to-Hematopoietic Transition. Homo sapiens

In order to dissect Endothelial-to-Hematopoietic Transition (EHT) in human iPS differentiated cultures, at day ten of differentiation towards the hematopoietic lineage, CD34+ single cells were sorted and analysed by single-cell qRT-PCR for 91 genes selected from the publication record, related to endothelium and hematopoietic stem and progenitor cells, as well as to the EHT in mouse and human. Overall design: To ensure a good representation of both endothelial and hematopoietic fractions, each 96-well plate received equal numbers of CD43 +/- cells, using near overlapping gates. A total of 437 CD34+ single-cells sorted in 5 different plates were analysed. Following reverse transcription and pre-amplification, the samples were diluted 1:5 and run in Fluidigm 96:96 arrays on a Biomark device. 10 cell controls, no RT controls and no Template controls were run in parallel