Identification of novel genes and networks governing hematopoietic stem cell development

DOI: 10.15252/embr.201642395

Authors: Tianxu Han; Chao‐Shun Yang; Kung‐Yen Chang; Danhua Zhang; Farhad B Imam; Tariq M Rana

Step 1: iPSC/ESC maintainance

day 0 ~ 2

culture “adherent” mouse ESC in “DMEM” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.

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

Step 2: mesoderm 1

day 2 ~ 4

culture “spheroid” mouse ESC in “IMDM” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “ME” should express “CDH5”, “PECAM1”, “TEK”, “Scl”, “CD41”, “KDR”, “Cxcr4” at the end of culture.

Step 3: mesoderm 2

day 4 ~ 5 (duration missing in database; assumed 1 day)

culture “adherent” mouse ME in “IMDM”. “ME” should be “upregulated” for “HoxB4”. Medium should contain factors shown in the following table.