Engineering hiPSC-CM and hiPSC-EC laden 3D nanofibrous splenic hydrogel for improving cardiac function through revascularization and remuscularization in infarcted heart

DOI: 10.1016/j.bioactmat.2021.04.010

Authors: Ge Guan; Da Huo; Yanzhao Li; Xiaolin Zhao; Yinghao Li; Zhongliang Qin; Dayu Sun; Guanyuan Yang; Mingcan Yang; Ju Tan; Wen Zeng; Chuhong Zhu

Step 1: iPSC/ESC maintainance

day 0 ~ 1

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

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

Step 2: Primitive streak 1

day 1 ~ 3

culture “adherent” human PSC in “RPMI-1640” for “2” days. For ECM, use “SpGel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Step 3: Primitive streak 2

day 3 ~ 5

culture “adherent” human PSC in “RPMI-1640” for “2” days. For ECM, use “SpGel” 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 1

day 5 ~ 8

culture “adherent” human PSC in “RPMI-1640” for “3” days. For ECM, use “SpGel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Step 5: endothelial 1

day 8 ~ 15

culture “adherent” human PSC in “EGM2” for “7” days. For ECM, use “SpGel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “7” days.

Ideally, “EC” should express “PECAM1” at the end of culture.