Protocol for differentiating hematopoietic progenitor cells from human pluripotent stem cells in chemically defined monolayer culture

DOI: 10.1016/j.xpro.2024.103545

Authors: Shaokang Mo; Kengyuan Qu; Jun Shen; Kuangyu Yen

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human cell in “E8” 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, “dummy” should express “TRA-1-81”, “SSEA4” at the end of culture.

Step 2: mesoderm 1

day 1 ~ 3

culture “adherent” unknown species cell in “APEL 2 medium” for “2” days. For ECM, use “vitronectin” 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 “TBXT” at the end of culture.

Step 3: endothelial 1

day 3 ~ 4

culture “adherent” unknown species cell in “APEL 2 medium” for “1” days. For ECM, use “vitronectin” at “N/A” “N/A”. 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 4: endothelial 2

day 4 ~ 5

culture “adherent” unknown species cell in “APEL 2 medium” for “1” days. For ECM, use “vitronectin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “EC” should express “CD34”, “CDH5” at the end of culture.