Robust Revascularization in Models of Limb Ischemia Using a Clinically Translatable Human Stem Cell-Derived Endothelial Cell Product

DOI: 10.1016/j.ymthe.2018.03.017

Authors: Mark G. MacAskill; Jaimy Saif; Alison Condie; Maurits A. Jansen; Thomas J. MacGillivray; Adriana A.S. Tavares; Lucija Fleisinger; Helen L. Spencer; Marie Besnier; Ernesto Martin; Giovanni Biglino; David E. Newby; Patrick W.F. Hadoke; Joanne C. Mountford; Costanza Emanueli; Andrew H. Baker

Step 1: iPSC/ESC maintainance

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

culture “adherent” human ESC in “StemPro medium”. Medium should contain factors shown in the following table.

Step 2: Primitive streak 1

day 1 ~ 4

culture “adherent” human ESC in “N2B27 medium” for “3” days. For ECM, use “FN1” at “0.03” “g/m2”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Step 3: mesoderm 1

day 4 ~ 5

culture “adherent” human ESC in “StemPro” for “1” days. For ECM, use “FN1” at “0.03” “g/m2”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Step 4: endothelial 1

day 5 ~ 6

culture “adherent” human ESC in “EGM2” for “1” days. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “EC” should express “PECAM1”, “CDH5”, “KDR”, “CD34” at the end of culture. Ideally, “MSC” should be obtained at the end of culture. Ideally, “Pericyte/SMC” should be obtained at the end of culture.