Guiding T lymphopoiesis from pluripotent stem cells by defined transcription factors

DOI: 10.1038/s41422-019-0251-7

Authors: Rongqun Guo; Fangxiao Hu; Qitong Weng; Cui Lv; Hongling Wu; Lijuan Liu; Zongcheng Li; Yang Zeng; Zhijie Bai; Mengyun Zhang; Yuting Liu; Xiaofei Liu; Chengxiang Xia; Tongjie Wang; Peiqing Zhou; Kaitao Wang; Yong Dong; Yuxuan Luo; Xiangzhong Zhang; Yuxian Guan; Yang Geng; Juan Du; Yangqiu Li; Yu Lan; Jiekai Chen; Bing Liu; Jinyong Wang

Step 1: iPSC/ESC maintainance

day 0 ~ 1 (duration missing in database; assumed 1 day)

culture “adherent” mouse PSC in “DMEM”. “PSC” should be “upregulated” for “RUNX1”, “upregulated” for “Hoxa9”. Medium should contain factors shown in the following table.

Step 2: Primitive streak 1

day 1 ~ 3

culture “spheroid” mouse PSC in “IMDM” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Step 3: mesoderm 1

day 3 ~ 6

culture “adherent” mouse PSC in “IMDM” for “3” days. For ECM, use “Gelatin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Step 4: endothelial 1

day 6 ~ 8

culture “adherent” mouse PSC in “IMDM” for “2” days. “PSC” should be “upregulated” for “RUNX1”, “upregulated” for “Hoxa9”. For ECM, use “Gelatin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “EC” should express “CDH5”, “Esam”, “Tek”, “Fli1”, “Erg”, “Lmo2”, “Mycn”, “Sox7”, “Bcl11a”, “Hoxb5”, “Sox18”, “Procr”, “Cd47”, “Cd63”, “Nrp1”, “Efnb2”, “HEY1”, “Nrp2”, “Nr2f2”, “Ephb4”, “Ets1”, “Lyl1”, “Tal1”, “RUNX1”, “Hoxa9”, “GATA2” at the end of culture.