Pluripotent stem cell model of Shwachman–Diamond syndrome reveals apoptotic predisposition of hemoangiogenic progenitors

DOI: 10.1038/s41598-020-71844-8

Authors: Takayuki Hamabata; Katsutsugu Umeda; Kagehiro Kouzuki; Takayuki Tanaka; Tomoo Daifu; Seishiro Nodomi; Satoshi Saida; Itaru Kato; Shiro Baba; Hidefumi Hiramatsu; Mitsujiro Osawa; Akira Niwa; Megumu K. Saito; Yasuhiko Kamikubo; Souichi Adachi; Yoshiko Hashii; Akira Shimada; Hiroyoshi Watanabe; Kenji Osafune; Keisuke Okita; Tatsutoshi Nakahata; Kenichiro Watanabe; Junko Takita; Toshio Heike

Step 1: iPSC/ESC maintainance

day 0 ~ 7

culture “adherent” human PSC in “primate ESC medium” for “7” days. Medium should contain factors shown in the following table, and should be changed once in “7” days.

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

Step 2: mesoderm 1

day 7 ~ 13

culture “adherent” human PSC in “Stemline II” for “6” 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 “6” days.

Ideally, “mesoderm” should express “KDR”, “CD34” at the end of culture.

Step 3: endothelial 1

day 13 ~ 20

culture “adherent” human mesoderm in “alpha-MEM” for “7” days. Medium should contain factors shown in the following table, and should be changed once in “7” days.

Ideally, “EC” should express “PECAM1”, “CDH5”, “CD34”, “CD141”, “MCAM”, “KDR” at the end of culture.