Generation of vascularized pancreatic progenitors through co-differentiation of endoderm and mesoderm from human pluripotent stem cells

DOI: 10.1186/s13287-024-04120-5

Authors: Xiaopu Sang; Junming Xu; Yihang Wang; Jingyi Li; Jiasen Xu; Xiaoni Chen; Xianjie Shi; Fenfang Wu

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 be changed once in “1” days.

Ideally, “PSC” should express “NANOG”, “POU5F1” at the end of culture.

Step 2: Primitive streak 1

day 1 ~ 4

culture “adherent” human PSC in “MCDB131” for “3” 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 “3” days.

Ideally, “mesoderm” should express “TBXT”, “MIXL1”, “SFRP2”, “SMAD3” at the end of culture. Ideally, “endoderm” should express “SOX17”, “FOXA2” at the end of culture.

Step 3: mesoderm 1

day 4 ~ 6

culture “adherent” human PSC in “MCDB131” 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 6 ~ 11

culture “adherent” human PSC in “MCDB131” for “5” 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 “5” days.

Ideally, “EC” should express “CD34”, “PECAM1”, “CDH5” at the end of culture. Ideally, “PP” should express “PDX1”, “SOX9”, “NKX6.1” at the end of culture. Ideally, “Pericyte/SMC” should be obtained at the end of culture.