Development of human iPSC-derived quiescent hepatic stellate cell-like cells for drug discovery and in vitro disease modeling

DOI: 10.1016/j.stemcr.2021.11.002

Authors: Yuta Koui; Misao Himeno; Yusuke Mori; Yasuhiro Nakano; Eiko Saijou; Naoki Tanimizu; Yoshiko Kamiya; Hiroko Anzai; Natsuki Maeda; Luyao Wang; Tadanori Yamada; Yasuyuki Sakai; Ryuichiro Nakato; Atsushi Miyajima; Taketomo Kido

Step 1: iPSC/ESC maintainance

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

culture “adherent” human PSC in “StemFlex Medium”. For ECM, use “Vitronectin” at “N/A” “N/A”.

Ideally, “PSC” should express “OCT3/4” at the end of culture.

Step 2: Primitive streak 1

day 1 ~ 2

culture “spheroid” human PSC in “StemPro” for “1” days. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Step 3: Primitive streak 2

day 2 ~ 5

culture “spheroid” human PSC in “StemPro” for “3” days. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Step 4: mesoderm 1

day 5 ~ 9

culture “spheroid” human PSC in “StemPro” for “4” days. Medium should contain factors shown in the following table, and should be changed once in “4” days.

Ideally, “mesoderm” should express “MESP1”, “TBXT” at the end of culture.