Cells sorted off hiPSC-derived kidney organoids coupled with immortalized cells reliably model the proximal tubule

DOI: 10.1038/s42003-023-04862-7

Authors: Ramin Banan Sadeghian; Ryohei Ueno; Yuji Takata; Akihiko Kawakami; Cheng Ma; Toshikazu Araoka; Minoru Takasato; Ryuji Yokokawa

Step 1: iPSC/ESC maintainance

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

culture “adherent” human PSC in “StemFit”. For ECM, use “Laminin” at “0.003” “g/l”. 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 ~ 6

culture “adherent” human PSC in “APEL” for “5” days. For ECM, use “Laminin” at “0.003” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “5” days.

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

Step 3: mesoderm 1

day 6 ~ 8

culture “adherent” human PS in “APEL” for “2” days. For ECM, use “Laminin” at “0.003” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

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

Step 4: mesoderm 2

day 8 ~ 13

culture “spheroid” human IM in “APEL” for “5” days. Medium should contain factors shown in the following table, and should be changed once in “5” days.

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