Efficient Differentiation of Human Pluripotent Stem Cells to Endothelial Progenitors via Small-Molecule Activation of WNT Signaling

DOI: 10.1016/j.stemcr.2014.09.005

Authors: Xiaojun Lian; Xiaoping Bao; Abraham Al-Ahmad; Jialu Liu; Yue Wu; Wentao Dong; Kaitlin K. Dunn; Eric V. Shusta; Sean P. Palecek

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “mTeSR” 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, “PSC” should express “POU5F1” at the end of culture.

Step 2: Primitive streak 1

day 1 ~ 3

culture “adherent” human PSC in “DMEM/F12” 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, “PS” should express “brachyury” at the end of culture.

Step 3: endothelial 1

day 3 ~ 6

culture “adherent” human PS 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, “EPC” should express “CD34”, “PECAM1”, “CDH5”, “CD117”, “KDR”, “TIE-2” at the end of culture.

Step 4: endothelial 2

day 6 ~ 7 (duration missing in database; assumed 1 day)

culture “adherent” human EPC in “EGM2”. For ECM, use “Collagen” at “N/A” “N/A”.

Ideally, “EC” should express “PECAM1”, “CDH5”, “VWF”, “ICAM1”, “KDR” at the end of culture.