RNAseq of hPSC derived hemogenic endothelium and natural killer cells

Human pluripotent stem cell (hPSC)-derived hemogenic endothelium (HE) was specified using the following protocol: https://doi.org/10.1016/j.ymeth.2015.10.001. Specifically, WNT-independent (WNTi-) and WNT-dependent (WNTd-) specified HE was isolated as CD34+CD184-CD73- by FACS on Day 6 and 8, respectively in biological triplicate. hPSC-derived NK cells were derived by isolation of WNTi CD34+CD43+ hematopoietic progenitors and WNTd CD34+CD43-CD73-CD184- HE using FACS on Day 8 of hematopoietic differentiation. CD34+ cells were also isolated from cord blood samples in triplicate via FACS. These populations were differentiated into NK cells, grown on OP9-DL4 stroma, in the presence of with IL-3 (first 5 days of culture), IL-7, FLT3-L, SCF, and IL-15 in biological triplicate. NK cells were isolated as DAPI-GFP-CD56+ by FACS after 26-28 days of OP9-DL4 co-culture. RNAseq was purified using the Qiagen miRNeasy kit and sequenced on a Ilumina HiSeq 2500.