Human iPS cell-engineered cardiac tissue sheets with cardiomyocytes and vascular cells for cardiac regeneration

DOI: 10.1038/srep06716

Authors: Hidetoshi Masumoto; Takeshi Ikuno; Masafumi Takeda; Hiroyuki Fukushima; Akira Marui; Shiori Katayama; Tatsuya Shimizu; Tadashi Ikeda; Teruo Okano; Ryuzo Sakata; Jun K. Yamashita

Step 1: iPSC/ESC maintainance

day 0 ~ 1 (duration missing in database; assumed 1 day)

culture “adherent” human PSC in “MEF-CM”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table.

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

Step 2: Primitive streak 1

day 1 ~ 2

culture “adherent” human PSC in “RPMI1640” for “1” 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 “1” days.

Step 3: mesoderm 1

day 2 ~ 6

culture “adherent” unknown species cell and human mesoderm and human PSC in “RPMI1640” for “4” 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 “4” days.

Ideally, “mesoderm” should express “KDR”, “ISL1” at the end of culture.

Step 4: endothelial 1

day 6 ~ 8

culture “adherent” unknown species cell and human mesoderm 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.

Ideally, “EC” should express “CDH5” at the end of culture. Ideally, “CM” should express “cTnT” at the end of culture. Ideally, “MC” should express “PDGFRb” at the end of culture.