Isolation of primitive endoderm, mesoderm, vascular endothelial and trophoblast progenitors from human pluripotent stem cells

DOI: 10.1038/nbt.2239

Authors: Micha Drukker; Chad Tang; Reza Ardehali; Yuval Rinkevich; Jun Seita; Andrew S Lee; Adriane R Mosley; Irving L Weissman; Yoav Soen

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”.

Step 2: Primitive streak 1

day 1 ~ 4

culture “spheroid” human PSC in “DMEM/F12” for “3” 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 “3” days.

Ideally, “PS” should express “TBXT”, “MESP1” at the end of culture.

Step 3: mesoderm 1

day 4 ~ 11

culture “adherent” human PSC in “DMEM/F12” for “7” 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 “7” days.

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

Step 4: endothelial 1

day 11 ~ 12 (duration missing in database; assumed 1 day)

culture “unspecified method” unknown species cell in “unspecified medium”.

Ideally, “EC” should express “VCAM1”, “VEGFC” at the end of culture.