Disease Model of GATA4 Mutation Reveals Transcription Factor Cooperativity in Human Cardiogenesis

DOI: 10.1016/j.cell.2016.11.033

Authors: Yen-Sin Ang; Renee N. Rivas; Alexandre J.S. Ribeiro; Rohith Srivas; Janell Rivera; Nicole R. Stone; Karishma Pratt; Tamer M.A. Mohamed; Ji-Dong Fu; C. Ian Spencer; Nathaniel D. Tippens; Molong Li; Anil Narasimha; Ethan Radzinsky; Anita J. Moon-Grady; Haiyuan Yu; Beth L. Pruitt; Michael P. Snyder; Deepak Srivastava

Step 1: iPSC/ESC maintainance

day 0 ~ 3

culture “adherent” human PSC in “mTeSR” 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, “PSC” should express “TRA-1-60”, “TRA-1-81”, “NANOG”, “POU5F1”, “SSEA4” at the end of culture.

Step 2: Primitive streak 1

day 3 ~ 5

culture “adherent” human PSC in “RPMI1640” 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: mesoderm 1

day 5 ~ 7

culture “adherent” unknown species cell in “RPMI1640” 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 4: mesoderm 2

day 7 ~ 9

culture “adherent” unknown species cell in “RPMI1640” for “2” days. For ECM, use “FN1” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “CPC” should express “ISL1”, “GATA4” at the end of culture.