Orthogonally induced differentiation of stem cells for the programmatic patterning of vascularized organoids and bioprinted tissues

DOI: 10.1038/s41551-022-00856-8

Authors: Mark A. Skylar-Scott; Jeremy Y. Huang; Aric Lu; Alex H. M. Ng; Tomoya Duenki; Songlei Liu; Lucy L. Nam; Sarita Damaraju; George M. Church; Jennifer A. Lewis

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “mTeSR” for “1” 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 “1” days.

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

Step 2: Primitive streak 1

day 1 ~ 3

culture “adherent” human PSC in “RPMI 1640” 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.

Step 3: mesoderm 1

day 3 ~ 5

culture “adherent” human PSC in “RPMI 1640” 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.

Step 4: endothelial 1

day 5 ~ 7

culture “adherent” human PSC in “neural induction medium” for “2” days. “PSC” should be “upregulated” for “ETV2”. 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 “CDH5”, “VWF”, “PECAM1”, “NRP1” at the end of culture.

Step 5: endothelial 2

day 7 ~ 9

culture “adherent” human PSC in “EGM2” 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 be obtained at the end of culture.