A Multi-Lineage Screen Reveals mTORC1 Inhibition Enhances Human Pluripotent Stem Cell Mesendoderm and Blood Progenitor Production

DOI: 10.1016/j.stemcr.2016.04.003

Authors: Emanuel Joseph Paul Nazareth; Nafees Rahman; Ting Yin; Peter William Zandstra

Step 1: iPSC/ESC maintainance

day 0 ~ 2

culture “adherent” human PS in “NutriStem” for “2” days. For ECM, use “Geltrex” at “2” “%”. Medium should be changed once in “2” days.

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

Step 2: Primitive streak 1

day 2 ~ 3 (duration missing in database; assumed 1 day)

culture “unspecified method” human PS in “unspecified medium”. Medium should contain factors shown in the following table.

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

Step 3: mesoderm 1

day 3 ~ 5

culture “spheroid” human PS in “StemPro” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “mesoderm” should express “BRACHYURY”, “MIXL1” at the end of culture.

Step 4: endothelial 1

day 5 ~ 11

culture “spheroid” human mesoderm in “StemPro” for “6” days. Medium should contain factors shown in the following table, and should be changed once in “6” days.

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