Bioprinting of human pluripotent stem cell derived corneal endothelial cells with hydrazone crosslinked hyaluronic acid bioink

DOI: 10.1186/s13287-024-03672-w

Authors: Pyry Grönroos; Anni Mörö; Paula Puistola; Karoliina Hopia; Maija Huuskonen; Taina Viheriälä; Tanja Ilmarinen; Heli Skottman

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “E8” for “1” days. For ECM, use “Laminin-521” at “N/A” “N/A”. Medium should be changed once in “1” days.

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

Step 2: endothelial 1

day 1 ~ 7

culture “adherent” human PSC in “KO-DMEM” for “6” days. For ECM, use “Laminin-521” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “6” days.

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

Step 3: endothelial 2

day 7 ~ 10

culture “adherent” human PSC in “KO-DMEM” for “3” days. For ECM, use “Laminin-521” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

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