Laminin-guided highly efficient endothelial commitment from human pluripotent stem cells

DOI: 10.1038/srep35680

Authors: Ryo Ohta; Akira Niwa; Yukimasa Taniguchi; Naoya M. Suzuki; Junko Toga; Emiko Yagi; Norikazu Saiki; Yoko Nishinaka-Arai; Chihiro Okada; Akira Watanabe; Tatsutoshi Nakahata; Kiyotoshi Sekiguchi; Megumu K. Saito

Step 1: iPSC/ESC maintainance

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

culture “adherent” human PSC in “mTeSR”. For ECM, use “LM511-E8” at “0.004” “g/m2”.

Step 2: mesoderm 1

day 1 ~ 3

culture “adherent” human PSC in “E8” for “2” days. For ECM, use “LM511-E8” at “0.004” “g/m2”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

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

Step 3: endothelial 1

day 3 ~ 7

culture “adherent” human mesoderm in “Stempro34 SFM” for “4” days. For ECM, use “LM411-E8” at “0.004” “g/m2”. Medium should contain factors shown in the following table, and should be changed once in “4” days.

Ideally, “EPC” should express “KDR”, “CD34”, “CDH5”, “CDH5”, “PECAM1”, “MCAM”, “LMO2”, “NOTCH1”, “HEY2”, “DLL4”, “NRP1”, “VWF”, “NOS3”, “IL8”, “IL6”, “IL18”, “NFKB1”, “EDN1”, “CAV1” at the end of culture.

Step 4: endothelial 2

day 7 ~ 10

culture “adherent” human EPC in “Stempro34 SFM” for “3” days. For ECM, use “LM411-E8” at “0.004” “g/m2”. Medium should be changed once in “3” days.

Ideally, “EC” should express “PECAM1”, “MCAM”, “CD143”, “Tie-2”, “EphB4” at the end of culture.