Expandable hESC-derived cardiovascular progenitor cells generate functional cardiac lineage cells for microtissue construction

DOI: 10.1186/s13287-024-03919-6

Authors: Siamak Rezaeiani; Malihe Rezaee; Mojtaba Shafaghi; Mohammad Karami; Roghayeh Hamidi; Hamid Khodayari; Sadaf Vahdat; Sara Pahlavan; Hossein Baharvand

Step 1: iPSC/ESC maintainance

day 0 ~ 5

culture “spheroid” human ESC in “unspecified medium” for “5” days. Medium should be changed once in “5” days.

Step 2: mesoderm 1

day 5 ~ 6

culture “spheroid” human ESC in “unspecified medium” for “1” days. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Step 3: mesoderm 2

day 6 ~ 8

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

Ideally, “CPC” should express “NKX2.5”, “ADGRL2”, “ISL1” at the end of culture.

Step 4: endothelial 1

day 8 ~ 10

culture “adherent” human CPC in “DMEM/F12” for “2” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

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