Renal Subcapsular Transplantation of PSC-Derived Kidney Organoids Induces Neo-vasculogenesis and Significant Glomerular and Tubular Maturation In Vivo

DOI: 10.1016/j.stemcr.2018.01.041

Authors: Cathelijne W. van den Berg; Laila Ritsma; M. Cristina Avramut; Loes E. Wiersma; Bernard M. van den Berg; Daniëlle G. Leuning; Ellen Lievers; Marije Koning; Jessica M. Vanslambrouck; Abraham J. Koster; Sara E. Howden; Minoru Takasato; Melissa H. Little; Ton J. Rabelink

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “E8” for “1” days. For ECM, use “vitronectin” 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 ~ 5

culture “adherent” human PSC in “STEMdiff APEL medium” for “4” days. For ECM, use “vitronectin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “4” days.

Step 3: mesoderm 1

day 5 ~ 8

culture “adherent” human PSC in “STEMdiff APEL medium” for “3” days. For ECM, use “vitronectin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Step 4: mesoderm 2

day 8 ~ 9 (duration missing in database; assumed 1 day)

culture “adherent” human PSC in “STEMdiff APEL medium”. For ECM, use “vitronectin” at “N/A” “N/A”. Medium should contain factors shown in the following table.

Step 5: endothelial 1

day 9 ~ 14

culture “device” human PSC in “STEMdiff APEL medium” for “5” days. Medium should contain factors shown in the following table, and should be changed once in “5” days.

Step 6: endothelial 2

day 14 ~ 16

culture “device” human PSC in “STEMdiff APEL medium” for “2” days. Medium should be changed once in “2” days.

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