Vasculogenesis in kidney organoids upon transplantation

DOI: 10.1038/s41536-022-00237-4

Authors: Marije Koning; Sébastien J. Dumas; M. Cristina Avramut; Roman I. Koning; Elda Meta; Ellen Lievers; Loes E. Wiersma; Mila Borri; Xue Liang; Lin Xie; Ping Liu; Fang Chen; Lin Lin; Yonglun Luo; Jaap Mulder; H. Siebe Spijker; Thierry Jaffredo; Bernard M. van den Berg; Peter Carmeliet; Cathelijne W. van den Berg; Ton J. Rabelink

Step 1: iPSC/ESC maintainance

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

culture “adherent” human PSC in “E8”. For ECM, use “Vitronectin” at “N/A” “N/A”. Medium should contain factors shown in the following table.

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

Step 2: Primitive streak 1

day 1 ~ 5

culture “adherent” human PSC in “APEL2 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” unknown species cell in “APEL2 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” unknown species cell in “APEL2 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” unknown species cell in “APEL2 medium” for “5” days. Medium should contain factors shown in the following table, and should be changed once in “5” days.

Ideally, “EC” should express “PECAM1”, “ANGPT2”, “PDGFB”, “TEK” at the end of culture.

Step 6: endothelial 2

day 14 ~ 16

culture “device” unknown species cell in “APEL2 medium” for “2” days. Medium should be changed once in “2” days.