Efficient Direct Reprogramming of Mature Amniotic Cells into Endothelial Cells by ETS Factors and TGFβ Suppression

DOI: 10.1016/j.cell.2012.09.032

Authors: Michael Ginsberg; Daylon James; Bi-Sen Ding; Daniel Nolan; Fuqiang Geng; Jason M. Butler; William Schachterle; Venkat R. Pulijaal; Susan Mathew; Stephen T. Chasen; Jenny Xiang; Zev Rosenwaks; Koji Shido; Olivier Elemento; Sina Y. Rabbany; Shahin Rafii

Step 1: iPSC/ESC maintainance

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

culture “adherent” human ESC in “KOSR medium”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table.

Step 2: mesoderm 1

day 1 ~ 11

culture “unspecified method” human ESC in “unspecified medium” for “10” days. Medium should contain factors shown in the following table, and should be changed once in “10” days.

Ideally, “mesoderm” should express “KDR” at the end of culture.

Step 3: endothelial 1

day 11 ~ 25

culture “adherent” human AC in “Medium 199” for “14” days. “AC” should be “upregulated” for “ETV2”, “upregulated” for “FLI1”, “upregulated” for “ERG1”. For ECM, use “FN1” at “0.001” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “14” days.

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

Step 4: endothelial 2

day 25 ~ 53

culture “adherent” human AC in “Medium 199” for “28” days. “AC” should be “downregulated” for “ETV2”, “upregulated” for “FLI1”, “upregulated” for “ERG1”. For ECM, use “FN1” at “0.001” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “28” days.

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