High-efficient serum-free differentiation of endothelial cells from human iPS cells

DOI: 10.1186/s13287-022-02924-x

Authors: Sarkawt Hamad; Daniel Derichsweiler; John Antonydas Gaspar; Konrad Brockmeier; Jürgen Hescheler; Agapios Sachinidis; Kurt Paul Pfannkuche

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “E8” for “1” days. For ECM, use “Matrigel” at “0.1” “g/m2”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

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

Step 2: Primitive streak 1

day 1 ~ 2 (duration missing in database; assumed 1 day)

culture “unspecified method” human PSC in “unspecified medium”.

Step 3: Primitive streak 2

day 2 ~ 3 (duration missing in database; assumed 1 day)

culture “unspecified method” human PSC in “unspecified medium”.

Step 4: mesoderm 1

day 3 ~ 4

culture “adherent” human PSC in “E6 medium” for “1” days. For ECM, use “Matrigel” at “0.1” “g/m2”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

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

Step 5: mesoderm 2

day 4 ~ 5

culture “adherent” human PSC in “E6 medium” for “1” days. For ECM, use “Matrigel” at “0.1” “g/m2”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

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

Step 6: endothelial 1

day 5 ~ 7

culture “adherent” human PSC in “E6 medium” for “2” days. For ECM, use “Matrigel” at “0.1” “g/m2”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “EC” should express “NOS3” at the end of culture.

Step 7: endothelial 2

day 7 ~ 9

culture “adherent” human PSC in “E6 medium” for “2” days. For ECM, use “Matrigel” at “0.1” “g/m2”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “EC” should express “NOS3”, “PECAM1”, “CDH5”, “VWF”, “CD34”, “CD184”, “LDL receptor” at the end of culture.