Biphasic modulation of insulin signaling enables highly efficient hematopoietic differentiation from human pluripotent stem cells

DOI: 10.1186/s13287-018-0934-x

Authors: Fuyu Duan; Rujin Huang; Fengzhi Zhang; Yonglin Zhu; Lin Wang; Xia Chen; Lufeng Bai; Wei Guo; Sophia Chia-Ning Chang; Xiaoyu Hu; Jie Na

Step 1: iPSC/ESC maintainance

day 0 ~ 2

culture “adherent” human PSC in “E8” 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, “PSC” should be obtained at the end of culture.

Step 2: mesoderm 1

day 2 ~ 3

culture “adherent” human PSC in “RPMI 1640” 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 be obtained at the end of culture.

Step 3: mesoderm 2

day 3 ~ 5

culture “adherent” human PSC in “RPMI 1640” 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 be obtained at the end of culture.

Step 4: endothelial 1

day 5 ~ 7

culture “adherent” human mesoderm in “RPMI 1640” 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 be obtained at the end of culture.

Step 5: endothelial 2

day 7 ~ 10

culture “adherent” human EC in “RPMI 1640” 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.

Ideally, “EC” should be obtained at the end of culture.