Improving hematopoietic differentiation from human induced pluripotent stem cells by the modulation of Hippo signaling with a diarylheptanoid derivative

DOI: 10.1186/s13287-024-03686-4

Authors: Umnuaychoke Thongsa-ad; Anongnat Wongpan; Wasinee Wongkummool; Phaewa Chaiwijit; Kwanchanok Uppakara; Gorawin Chaiyakitpattana; Passanan Singpant; Pirut Tong-ngam; Amnat Chukhan; Wachirachai Pabuprappap; Sirapope Wongniam; Apichart Suksamrarn; Suradej Hongeng; Usanarat Anurathapan; Kasem Kulkeaw; Alisa Tubsuwan; Kanit Bhukhai

Step 1: iPSC/ESC maintainance

day 0 ~ 1

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

Step 2: mesoderm 1

day 1 ~ 3

culture “adherent” human PSC in “STEMdiff APEL2” 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.

Step 3: endothelial 1

day 3 ~ 6

culture “adherent” human PSC in “STEMdiff APEL2” 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 express “CDH5”, “CD73” at the end of culture.

Step 4: endothelial 2

day 6 ~ 11

culture “adherent” human PSC in “STEMdiff APEL2” for “5” 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 “5” days.

Ideally, “EC” should express “CDH5”, “CD73” at the end of culture. Ideally, “HSPC” should express “CD34”, “PTPRC”, “THY1”, “WWTR1”, “TEAD1”, “TEAD3”, “YAP1”, “CYR61”, “CTGF”, “CCDN1”, “HEG1”, “RASIP1”, “RHOC”, “THY1”, “BMP4”, “WNT5A”, “SOX2”, “SNAI2”, “BMP7”, “TGFB2”, “WNT3”, “ARID5B”, “BCAR1”, “CDH2”, “COL3A1”, “COL5A1”, “HEY1”, “IGFBP3”, “TM4SF1”, “GLI3”, “cMYC” at the end of culture.