A Scalable and Efficient Bioprocess for Manufacturing Human Pluripotent Stem Cell-Derived Endothelial Cells

DOI: 10.1016/j.stemcr.2018.07.001

Authors: Haishuang Lin; Qian Du; Qiang Li; Ou Wang; Zhanqi Wang; Neety Sahu; Christian Elowsky; Kan Liu; Chi Zhang; Soonkyu Chung; Bin Duan; Yuguo Lei

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “spheroid” human PSC in “E8” for “1” days. For ECM, use “PNIPAAm-PEG hydrogel” at “10” “%”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “PSC” should express “OCT3/4”, “NANOG” at the end of culture.

Step 2: endothelial 1

day 1 ~ 4

culture “spheroid” human PSC in “N2B27 medium” for “3” days. For ECM, use “PNIPAAm-PEG hydrogel” at “10” “%”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Step 3: endothelial 2

day 4 ~ 5

culture “spheroid” human PSC in “StemPro-34 SFM medium” for “1” days. For ECM, use “PNIPAAm-PEG hydrogel” at “10” “%”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “EC” should express “PECAM1”, “CDH5” at the end of culture.