Integrative epigenomic and transcriptomic analysis reveals the requirement of JUNB for hematopoietic fate induction

DOI: 10.1038/s41467-022-30789-4

Authors: Xia Chen; Peiliang Wang; Hui Qiu; Yonglin Zhu; Xingwu Zhang; Yaxuan Zhang; Fuyu Duan; Shuangyuan Ding; Jianying Guo; Yue Huang; Jie Na

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “E8” for “1” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “PSC” should express “POU5F1”, “NANOG”, “SOX2” at the end of culture.

Step 2: mesoderm 1

day 1 ~ 2

culture “adherent” human PSC in “RPMI1640” for “1” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “mesoderm” should express “KDR”, “MIXL1”, “GATA6”, “HAND1”, “BMP2”, “COL3A1” at the end of culture.

Step 3: mesoderm 2

day 2 ~ 4

culture “adherent” human mesoderm in “RPMI1640” for “2” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Step 4: endothelial 1

day 4 ~ 6

culture “adherent” human mesoderm in “RPMI1640” for “2” days. For ECM, use “Matrigel” 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 “PECAM1”, “CD34”, “CDH5”, “ETV2”, “TAL1”, “SOX17”, “FLI1”, “KDR”, “EFNB2”, “GJA4”, “JUNB” at the end of culture.

Step 5: endothelial 2

day 6 ~ 9

culture “adherent” human EC in “RPMI1640” for “3” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “3” days.