Mapping human pluripotent stem cell differentiation pathways using high throughput single-cell RNA-sequencing

DOI: 10.1186/s13059-018-1426-0

Authors: Xiaoping Han; Haide Chen; Daosheng Huang; Huidong Chen; Lijiang Fei; Chen Cheng; He Huang; Guo-Cheng Yuan; Guoji Guo

Step 1: iPSC/ESC maintainance

day 0 ~ 20

culture “unspecified method” human ESC in “RSeT medium” for “20” days. Medium should contain factors shown in the following table, and should be changed once in “20” days.

Ideally, “ESC” should express “POU5F1”, “NANOG”, “PRDM14”, “NODAL”, “LEFTY2”, “GDF3”, “KLF4”, “DNMT3L”, “IL6ST”, “DPPA2”, “TDGF1”, “SOX2” at the end of culture.

Step 2: endothelial 1

day 20 ~ 28

culture “spheroid” unknown species cell and human ESC in “DMEM/F12” for “8” days. Medium should contain factors shown in the following table, and should be changed once in “8” days.

Ideally, “EC” should express “KDR”, “GNG11”, “ECSCR”, “CD34”, “PROCR”, “GATA2”, “TAL1”, “HOXA1”, “TFAP2A” at the end of culture.