Mitophagy mediates metabolic reprogramming of induced pluripotent stem cells undergoing endothelial differentiation

DOI: 10.1016/j.jbc.2021.101410

Authors: Sarah Krantz; Young-Mee Kim; Shubhi Srivastava; Joseph W. Leasure; Peter T. Toth; Glenn Marsboom; Jalees Rehman

Step 1: iPSC/ESC maintainance

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

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

Step 2: mesoderm 1

day 1 ~ 2

culture “adherent” human PSC in “IMDM” 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.

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

Step 3: mesoderm 2

day 2 ~ 3

culture “adherent” human PSC in “IMDM” 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 4: endothelial 1

day 3 ~ 4

culture “adherent” human mesoderm in “IMDM” 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.

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

Step 5: endothelial 2

day 4 ~ 5 (duration missing in database; assumed 1 day)

culture “adherent” human EC in “EGM2-MV”. For ECM, use “Gelatin” at “0.2” “%”. Medium should contain factors shown in the following table.

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