Vitamin C–dependent lysine demethylase 6 (KDM6)-mediated demethylation promotes a chromatin state that supports the endothelial-to-hematopoietic transition

DOI: 10.1074/jbc.RA119.009757

Authors: Tian Zhang; Ke Huang; Yanling Zhu; Tianyu Wang; Yongli Shan; Bing Long; Yuhang Li; Qianyu Chen; Pengtao Wang; Shaoyang Zhao; Dongwei Li; Chuman Wu; Baoqiang Kang; Jiaming Gu; Yuchan Mai; Qing Wang; Jinbing Li; Yanqi Zhang; Zechuan Liang; Lin Guo; Fang Wu; Shuquan Su; Junwei Wang; Minghui Gao; Xiaofen Zhong; Baojian Liao; Jiekai Chen; Xiao Zhang; Xiaodong Shu; Duanqing Pei; Jinfu Nie; Guangjin Pan

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “mTeSR” 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.

Step 2: Primitive streak 1

day 1 ~ 2

culture “adherent” human PSC in “DMEM/F12” 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, “PS” should express “TBXT” at the end of culture.

Step 3: mesoderm 1

day 2 ~ 3

culture “adherent” human PSC in “DMEM/F12” 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, “LM” should express “HAND1”, “MSGN1” at the end of culture.

Step 4: endothelial 1

day 3 ~ 4

culture “adherent” human PSC in “DMEM/F12” 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, “HEC” should express “GATA2”, “PECAM1”, “RUNX1”, “MYB” at the end of culture.

Step 5: endothelial 2

day 4 ~ 5

culture “adherent” human PSC in “DMEM/F12” 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.