PRC2 specifies ectoderm lineages and maintains pluripotency in primed but not naïve ESCs

DOI: 10.1038/s41467-017-00668-4

Authors: Yongli Shan; Zechuan Liang; Qi Xing; Tian Zhang; Bo Wang; Shulan Tian; Wenhao Huang; Yanqi Zhang; Jiao Yao; Yanling Zhu; Ke Huang; Yujian Liu; Xiaoshan Wang; Qianyu Chen; Jian Zhang; Bizhi Shang; Shengbiao Li; Xi Shi; Baojian Liao; Cong Zhang; Keyu Lai; Xiaofen Zhong; Xiaodong Shu; Jinyong Wang; Hongjie Yao; Jiekai Chen; Duanqing Pei; Guangjin Pan

Step 1: iPSC/ESC maintainance

day 0 ~ 20

culture “adherent” human ESC and human ESC and human ESC and human ESC and human ESC and human ESC and human ESC and human ESC and human ESC and human ESC in “mTeSR” for “20” days. “ESC” should be “unspecified regulation” for “EED”, “unspecified regulation” for “SUZ12”, “unspecified regulation” for “EZH1”, “unspecified regulation” for “EZH2”. “ESC” should be “unspecified regulation” for “EZH1”, “unspecified regulation” for “EZH2”. “ESC” should be “downregulated” for “EZH2”, “downregulated” for “EZH1”. “ESC” should be “downregulated” for “EED”. “ESC” should be “downregulated” for “SUZ12”. “ESC” should be “upregulated” for “EED”. “ESC” should be “upregulated” for “SUZ12”. “ESC” should be “upregulated” for “NANOG”, “upregulated” for “KLF2”. “ESC” should be “upregulated” for “EZH2”. “ESC” should be “upregulated” for “EZH1”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “20” days.

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

Step 2: mesoderm 1

day 20 ~ 22

culture “adherent” human ESC in “E6 medium” 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, “mesoderm” should express “CALPONIN”, “CD34” at the end of culture.

Step 3: mesoderm 2

day 22 ~ 24

culture “adherent” human ESC in “E6 medium” 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.