Transdifferentiated Human Vascular Smooth Muscle Cells are a New Potential Cell Source for Endothelial Regeneration

DOI: 10.1038/s41598-017-05665-7

Authors: Xuechong Hong; Andriana Margariti; Alexandra Le Bras; Laureen Jacquet; Wei Kong; Yanhua Hu; Qingbo Xu

Step 1: Primitive streak 1

day 0 ~ 2

culture “adherent” human Pericyte/SMC in “KnockOut DMEM/F12” for “2” days. “Pericyte/SMC” should be “upregulated” for “POU5F1”, “upregulated” for “SOX2”, “upregulated” for “KLF4”, “upregulated” for “c-MYC”. For ECM, use “Gelatin” at “0.05” “%”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Step 2: endothelial 1

day 2 ~ 8

culture “adherent” human Pericyte/SMC in “EGM2” for “6” days. “Pericyte/SMC” should be “downregulated” for “HES5”, “upregulated” for “HES5”, “upregulated” for “JAG1”, “downregulated” for “JAG1”. For ECM, use “Collagen IV” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “6” days.

Ideally, “EC” should be obtained at the end of culture.

Step 3: endothelial 2

day 8 ~ 18

culture “adherent” human Pericyte/SMC in “EGM2” for “10” days. For ECM, use “Collagen IV” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “10” days.

Ideally, “EC” should be obtained at the end of culture.