Tissue-engineered blood-brain barrier models via directed differentiation of human induced pluripotent stem cells

DOI: 10.1038/s41598-019-50193-1

Authors: Gabrielle N. Grifno; Alanna M. Farrell; Raleigh M. Linville; Diego Arevalo; Joo Ho Kim; Luo Gu; Peter C. Searson

Step 1: iPSC/ESC maintainance

day 0 ~ 3

culture “adherent” human PSC and human PSC in “mTeSR” 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.

Step 2: Primitive streak 1

day 3 ~ 4

culture “adherent” human PSC and human PSC in “DMEM/F12” 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.

Step 3: mesoderm 1

day 4 ~ 5

culture “adherent” human PSC and human PSC in “DMEM/F12” 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, “mesoderm” should be obtained at the end of culture.

Step 4: endothelial 1

day 5 ~ 7

culture “adherent” human PSC and human PSC in “hESFM” 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, “BMEC” should express “PECAM1”, “ZO-1”, “claudin-5”, “occludin”, “GLUT-1”, “Pgp”, “CDH5” at the end of culture.