GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm

DOI: 10.1016/j.stemcr.2019.07.009

Authors: Julio Castaño; Sergi Aranda; Clara Bueno; Fernando J. Calero-Nieto; Eva Mejia-Ramirez; Jose Luis Mosquera; Enrique Blanco; Xiaonan Wang; Cristina Prieto; Lorea Zabaleta; Elisabetta Mereu; Meritxell Rovira; Senda Jiménez-Delgado; Daniel R. Matson; Holger Heyn; Emery H. Bresnick; Berthold Göttgens; Luciano Di Croce; Pablo Menendez; Angel Raya; Alessandra Giorgetti

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “mTeSR” for “1” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should be changed once in “1” days.

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

Step 2: mesoderm 1

day 1 ~ 2

culture “spheroid” human PSC in “serum-free medium” for “1” days. “PSC” should be “upregulated” for “GATA2”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “mesoderm” should express “TBXT”, “MIXL1”, “GATA2” at the end of culture.

Step 3: endothelial 1

day 2 ~ 9

culture “spheroid” human PSC in “serum-free medium” for “7” days. “PSC” should be “upregulated” for “GATA2”. Medium should contain factors shown in the following table, and should be changed once in “7” days.

Ideally, “HEP” should express “RUNX1”, “SCL”, “GATA2” at the end of culture.

Step 4: endothelial 2

day 9 ~ 14

culture “spheroid” human PSC in “serum-free medium” for “5” days. Medium should be changed once in “5” days.

Ideally, “HPC” should express “GATA2”, “RUNX1”, “SCL” at the end of culture.