Generation, functional analysis and applications of isogenic three-dimensional self-aggregating cardiac microtissues from human pluripotent stem cells

DOI: 10.1038/s41596-021-00497-2

Authors: Giulia Campostrini; Viviana Meraviglia; Elisa Giacomelli; Ruben W. J. van Helden; Loukia Yiangou; Richard P. Davis; Milena Bellin; Valeria V. Orlova; Christine L. Mummery

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “E8” for “1” days. For ECM, use “Vitronectin” 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 ~ 4

culture “adherent” human PSC in “LI-BPEL” for “3” days. For ECM, use “Matrigel” at “0.833” “%”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

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

Step 3: endothelial 1

day 4 ~ 7

culture “adherent” human mesoderm in “LI-BPEL” for “3” days. For ECM, use “Matrigel” at “0.833” “%”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Ideally, “EC” should express “CD34” at the end of culture.

Step 4: endothelial 2

day 7 ~ 8

culture “adherent” human EC in “LI-BPEL” for “1” days. For ECM, use “FN1” at “0.005” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “EC” should express “PECAM1”, “CDH5” at the end of culture.