Overexpression of GATA2 Enhances Development and Maintenance of Human Embryonic Stem Cell-Derived Hematopoietic Stem Cell-like Progenitors

DOI: 10.1016/j.stemcr.2019.05.007

Authors: Ya Zhou; Yonggang Zhang; Bo Chen; Yong Dong; Yimeng Zhang; Bin Mao; Xu Pan; Mowen Lai; Yijin Chen; Guohui Bian; Qiongxiu Zhou; Tatsutoshi Nakahata; Jiaxi Zhou; Min Wu; Feng Ma

Step 1: iPSC/ESC maintainance

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

culture “adherent” human ESC in “mTeSR”. For ECM, use “Matrigel” at “N/A” “N/A”.

Ideally, “ESC” should express “SOX2”, “POU5F1”, “SSEA4” at the end of culture.

Step 2: Primitive streak 1

day 1 ~ 2

culture “adherent” unknown species cell in “unspecified medium” for “1” days. Medium should be changed once in “1” days.

Step 3: Primitive streak 2

day 2 ~ 3

culture “adherent” unknown species cell in “unspecified medium” for “1” days. Medium should be changed once in “1” days.

Step 4: mesoderm 1

day 3 ~ 4

culture “adherent” human ESC in “unspecified medium” for “1” days. “ESC” should be “upregulated” for “GATA2”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Step 5: mesoderm 2

day 4 ~ 5

culture “adherent” unknown species cell in “unspecified medium” for “1” days. Medium should be changed once in “1” days.

Step 6: endothelial 1

day 5 ~ 6

culture “adherent” human ESC in “unspecified medium” for “1” days. “ESC” should be “upregulated” for “GATA2”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

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

Step 7: endothelial 2

day 6 ~ 7

culture “adherent” unknown species cell in “unspecified medium” for “1” days. Medium should be changed once in “1” days.