Long-term engrafting multilineage hematopoietic cells differentiated from human induced pluripotent stem cells

DOI: 10.1038/s41587-024-02360-7

Authors: Elizabeth S. Ng; Gulcan Sarila; Jacky Y. Li; Hasindu S. Edirisinghe; Ritika Saxena; Shicheng Sun; Freya F. Bruveris; Tanya Labonne; Nerida Sleebs; Alexander Maytum; Raymond Y. Yow; Chantelle Inguanti; Ali Motazedian; Vincenzo Calvanese; Sandra Capellera-Garcia; Feiyang Ma; Hieu T. Nim; Mirana Ramialison; Constanze Bonifer; Hanna K. A. Mikkola; Edouard G. Stanley; Andrew G. Elefanty

Step 1: iPSC/ESC maintainance

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

culture “adherent” human PSC in “E8”. For ECM, use “Matrigel” at “N/A” “N/A”.

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

Step 2: mesoderm 1

day 1 ~ 3

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

Step 3: mesoderm 2

day 3 ~ 5

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

Step 4: endothelial 1

day 5 ~ 7

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

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

Step 5: endothelial 2

day 7 ~ 9

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