Generation of vascular endothelial and smooth muscle cells from human pluripotent stem cells

DOI: 10.1038/ncb3205

Authors: Christoph Patsch; Ludivine Challet-Meylan; Eva C. Thoma; Eduard Urich; Tobias Heckel; John F. O’Sullivan; Stephanie J. Grainger; Friedrich G. Kapp; Lin Sun; Klaus Christensen; Yulei Xia; Mary H. C. Florido; Wei He; Wei Pan; Michael Prummer; Curtis R. Warren; Roland Jakob-Roetne; Ulrich Certa; Ravi Jagasia; Per-Ola Freskgård; Isaac Adatto; Dorothee Kling; Paul Huang; Leonard I. Zon; Elliot L. Chaikof; Robert E. Gerszten; Martin Graf; Roberto Iacone; Chad A. Cowan

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.

Step 2: mesoderm 1

day 1 ~ 4

culture “adherent” human PSC in “N2B27 medium” 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, “mesoderm” should express “TBXT”, “MIXL1”, “EOMES”, “FGF4” at the end of culture.

Step 3: endothelial 1

day 4 ~ 5

culture “adherent” human PSC in “StemPro” 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.

Step 4: endothelial 2

day 5 ~ 7

culture “adherent” human PSC in “StemPro” for “2” days. For ECM, use “FN1” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “EC” should express “CDH5”, “KDR”, “PECAM1”, “CD34”, “CD105”, “VWF”, “PECAM1”, “SOX17” at the end of culture.