Multilineage differentiation potential of hematoendothelial progenitors derived from human induced pluripotent stem cells

DOI: 10.1186/s13287-020-01997-w

Authors: Ratchapong Netsrithong; Siriwal Suwanpitak; Bootsakorn Boonkaew; Kongtana Trakarnsanga; Lung-Ji Chang; Chartsiam Tipgomut; Chinnavuth Vatanashevanopakorn; Kovit Pattanapanyasat; Methichit Wattanapanitch

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 “2.5” “%”. Medium should contain factors shown in the following table.

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

Step 2: Primitive streak 1

day 1 ~ 2

culture “adherent” human PSC in “RPMI1640” for “1” days. For ECM, use “Matrigel” at “2.5” “%”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “PS” should express “TBXT” at the end of culture.

Step 3: mesoderm 1

day 2 ~ 3

culture “adherent” human PSC in “RPMI1640” for “1” days. For ECM, use “Matrigel” at “2.5” “%”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “HEP” should express “CDH5”, “CD34”, “PECAM1”, “KDR” at the end of culture.

Step 4: mesoderm 2

day 3 ~ 4 (duration missing in database; assumed 1 day)

culture “adherent” human PSC in “RPMI1640”. For ECM, use “Matrigel” at “2.5” “%”. Medium should contain factors shown in the following table.

Ideally, “HSPC” should express “CD34”, “CD43” at the end of culture.

Step 5: endothelial 1

day 4 ~ 6

culture “adherent” human HEP in “EGM” for “2” days. For ECM, use “Gelatin” at “0.1” “%”. Medium should be changed once in “2” days.

Ideally, “EC” should express “PECAM1”, “ICAM1”, “CDH5”, “VWF”, “CD34”, “KDR” at the end of culture.