Chemically-defined albumin-free differentiation of human pluripotent stem cells to endothelial progenitor cells

DOI: 10.1016/j.scr.2015.05.004

Authors: Xiaoping Bao; Xiaojun Lian; Kaitlin K. Dunn; Mengxuan Shi; Tianxiao Han; Tongcheng Qian; Vijesh J. Bhute; Scott G. Canfield; Sean P. Palecek

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “E8” for “1” days. For ECM, use “Synthemax” 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 be obtained at the end of culture.

Step 2: Primitive streak 1

day 1 ~ 3

culture “adherent” human PSC in “DMEM” for “2” days. For ECM, use “Synthemax” 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 “TBXT”, “MIXL1”, “EOMES” at the end of culture.

Step 3: mesoderm 1

day 3 ~ 6

culture “adherent” human PSC in “DMEM” for “3” days. For ECM, use “Synthemax” 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 “KDR”, “CD34”, “CDH5”, “PECAM1” at the end of culture.

Step 4: endothelial 1

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

culture “adherent” human EPC in “human endothelial-SFM”. For ECM, use “Collagen” at “N/A” “N/A”. Medium should contain factors shown in the following table.

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