Activation of lineage competence in hemogenic endothelium precedes the formation of hematopoietic stem cell heterogeneity

DOI: 10.1038/s41422-023-00797-0

Authors: Jun Xia; Mengyao Liu; Caiying Zhu; Shicheng Liu; Lanlan Ai; Dongyuan Ma; Ping Zhu; Lu Wang; Feng Liu

Step 1: iPSC/ESC maintainance

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

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

Step 2: Primitive streak 1

day 1 ~ 2 (duration missing in database; assumed 1 day)

culture “spheroid” human PSC in “unspecified medium”. Medium should contain factors shown in the following table.

Step 3: mesoderm 1

day 2 ~ 3 (duration missing in database; assumed 1 day)

culture “spheroid” human PSC in “unspecified medium”. Medium should contain factors shown in the following table.

Step 4: mesoderm 2

day 3 ~ 4 (duration missing in database; assumed 1 day)

culture “spheroid” human PSC in “unspecified medium”. Medium should contain factors shown in the following table.

Step 5: endothelial 1

day 4 ~ 5 (duration missing in database; assumed 1 day)

culture “spheroid” human PSC in “unspecified medium”. Medium should contain factors shown in the following table.

Step 6: endothelial 2

day 5 ~ 6 (duration missing in database; assumed 1 day)

culture “spheroid” human HEC in “unspecified medium”. “HEC” should be “downregulated” for “SPI1”. Medium should contain factors shown in the following table.

Ideally, “HEC” should express “DLL4”, “RUNX1”, “SPI1”, “CMYB”, “HLF”, “GFI1”, “CEBPB”, “LCP1”, “CSFIR” at the end of culture. Ideally, “HSPC” should express “DLL4”, “RUNX1”, “SPI1” at the end of culture.