Fetal sheep support the development of hematopoietic cells in vivo from human induced pluripotent stem cells

DOI: 10.1016/j.exphem.2020.12.006

Authors: Tomoyuki Abe; Hideki Uosaki; Hiroaki Shibata; Hiromasa Hara; Borjigin Sarentonglaga; Yoshikazu Nagao; Yutaka Hanazono

Step 1: iPSC/ESC maintainance

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

culture “adherent” human PSC in “Primate ES Cell Medium”. For ECM, use “MEF feeder” 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: mesoderm 1

day 1 ~ 2 (duration missing in database; assumed 1 day)

culture “adherent” human PSC in “unspecified medium”.

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

Step 3: mesoderm 2

day 2 ~ 3 (duration missing in database; assumed 1 day)

culture “adherent” human PSC in “unspecified medium”.

Ideally, “hemangioblast” should express “GATA2”, “ETV2”, “KDR”, “PDGFRA” at the end of culture.

Step 4: endothelial 1

day 3 ~ 5

culture “adherent” human PSC in “IMDM” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “HE” should express “CD34”, “KDR”, “PECAM1”, “CDH5”, “TIE2” at the end of culture.