Generation of kidney organoids from human pluripotent stem cells

DOI: 10.1038/nprot.2016.098

Authors: Minoru Takasato; Pei X Er; Han S Chiu; Melissa H Little

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “MEF-CM” 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, “PSC” should be obtained at the end of culture.

Step 2: Primitive streak 1

day 1 ~ 3

culture “adherent” human PSC in “APEL medium” 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”, “MIXL1” at the end of culture.

Step 3: mesoderm 1

day 3 ~ 5

culture “adherent” human PSC in “APEL medium” 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, “IM” should express “GATA3”, “HOXD11” at the end of culture.

Step 4: endothelial 1

day 5 ~ 7

culture “spheroid” human PSC in “APEL medium” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “EC” should express “PECAM1”, “KDR”, “SOX17” at the end of culture.

Step 5: endothelial 2

day 7 ~ 9

culture “spheroid” human PSC in “APEL medium” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.

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