The role of mutations associated with familial neurodegenerative disorders on blood–brain barrier function in an iPSC model

DOI: 10.1186/s12987-019-0139-4

Authors: Moriah E. Katt; Lakyn N. Mayo; Shannon E. Ellis; Vasiliki Mahairaki; Jeffrey D. Rothstein; Linzhao Cheng; Peter C. Searson

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human PSC in “E8” for “1” days. For ECM, use “Vitronectin” 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: mesoderm 1

day 1 ~ 6

culture “adherent” human PSC in “unspecified medium” for “5” days. For ECM, use “Vitronectin” at “N/A” “N/A”. Medium should be changed once in “5” days.

Step 3: endothelial 1

day 6 ~ 8

culture “adherent” unknown species cell in “unspecified medium” for “2” days. For ECM, use “Vitronectin” 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 “CLDN5”, “OCLN”, “ABCB1”, “SLC2A1” at the end of culture.

Step 4: endothelial 2

day 8 ~ 10

culture “adherent” unknown species cell in “unspecified medium” for “2” days. For ECM, use “Collagen IV and Fibronectin” at “N/A” “N/A”. Medium should be changed once in “2” days.

Ideally, “EC” should express “CLDN5”, “OCLN”, “ABCB1”, “SLC2A1” at the end of culture.