Nod1-dependent NF-kB activation initiates hematopoietic stem cell specification in response to small Rho GTPases

DOI: 10.1038/s41467-023-43349-1

Authors: Xiaoyi Cheng; Radwa Barakat; Giulia Pavani; Masuma Khatun Usha; Rodolfo Calderon; Elizabeth Snella; Abigail Gorden; Yudi Zhang; Paul Gadue; Deborah L. French; Karin S. Dorman; Antonella Fidanza; Clyde A. Campbell; Raquel Espin-Palazon

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “StemPro hESC SFM” for “1” days. For ECM, use “Vitronectin” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Step 2: Primitive streak 1

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

culture “spheroid” human PSC in “SFD medium”. Medium should contain factors shown in the following table.

Step 3: mesoderm 1

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

culture “spheroid” human PSC in “SFD medium”. Medium should contain factors shown in the following table.

Step 4: endothelial 1

day 3 ~ 8

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

Ideally, “EC” should be obtained at the end of culture. Ideally, “HE” should express “RUNX1” at the end of culture.