Assembly of Human Stem Cell-Derived Cortical Spheroids and Vascular Spheroids to Model 3-D Brain-like Tissues

DOI: 10.1038/s41598-019-42439-9

Authors: Liqing Song; Xuegang Yuan; Zachary Jones; Kyle Griffin; Yi Zhou; Teng Ma; Yan Li

Step 1: iPSC/ESC maintainance

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

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

Step 2: Primitive streak 1

day 1 ~ 4

culture “spheroid” unknown species cell and human PSC in “RPMI” for “3” days. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Step 3: mesoderm 1

day 4 ~ 9

culture “spheroid” unknown species cell and human PSC in “RPMI” for “5” days. Medium should contain factors shown in the following table, and should be changed once in “5” days.

Step 4: endothelial 1

day 9 ~ 15

culture “spheroid” unknown species cell and human PSC in “RPMI” for “6” days. Medium should contain factors shown in the following table, and should be changed once in “6” days.

Ideally, “EC” should express “PECAM1”, “CDH5”, “ZO1”, “GLUT-1”, “BCBP”, “Notch-1”, “KDR” at the end of culture.