Directed differentiation and long-term maintenance of epicardial cells derived from human pluripotent stem cells under fully defined conditions

DOI: 10.1038/nprot.2017.080

Authors: Xiaoping Bao; Xiaojun Lian; Tongcheng Qian; Vijesh J Bhute; Tianxiao Han; Sean P Palecek

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “E8” for “1” days. For ECM, use “Synthemax” at “0.025” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “PSC” should express “POU5F1” at the end of culture.

Step 2: Primitive streak 1

day 1 ~ 2

culture “adherent” human PSC in “RPMI” for “1” days. For ECM, use “Synthemax” at “0.025” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “PS” should express “TBXT” at the end of culture.

Step 3: mesoderm 1

day 2 ~ 4

culture “adherent” human PSC in “RPMI” for “2” days. For ECM, use “Synthemax” at “0.025” “g/l”. Medium should be changed once in “2” days.

Step 4: mesoderm 2

day 4 ~ 6

culture “adherent” human PSC in “RPMI” for “2” days. For ECM, use “Synthemax” at “0.025” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “2” days.