A combined hiPSC-derived endothelial cell and in vitro microfluidic platform for assessing biomaterial-based angiogenesis

DOI: 10.1016/j.biomaterials.2018.11.032

Authors: Sylvia L. Natividad-Diaz; Shane Browne; Amit K. Jha; Zhen Ma; Samir Hossainy; Yosuke K. Kurokawa; Steven C. George; Kevin E. Healy

Step 1: iPSC/ESC maintainance

day 0 ~ 2

culture “adherent” human PSC in “E8” 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, “PSC” should express “POU5F1”, “NANOG” at the end of culture.

Step 2: Primitive streak 1

day 2 ~ 4

culture “adherent” human PSC in “E8” 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 “TBXT” at the end of culture.

Step 3: mesoderm 1

day 4 ~ 5

culture “adherent” human PSC in “E8” 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, “EC” should express “CD34”, “PECAM1”, “CDH5”, “KDR”, “FLT1”, “ENG” at the end of culture. Ideally, “PSC” should express “POU5F1”, “NANOG” at the end of culture.

Step 4: endothelial 1

day 5 ~ 7

culture “adherent” human EC in “EGM2” for “2” days. For ECM, use “Gelatin” at “0.2” “%”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “EC” should express “PECAM1”, “NOS3”, “CDH5”, “VWF”, “KDR” at the end of culture.