Differential HDAC6 Activity Modulates Ciliogenesis and Subsequent Mechanosensing of Endothelial Cells Derived from Pluripotent Stem Cells

DOI: 10.1016/j.celrep.2018.06.083

Authors: Quinton Smith; Bria Macklin; Xin Yi Chan; Hannah Jones; Michelle Trempel; Mervin C. Yoder; Sharon Gerecht

Step 1: iPSC/ESC maintainance

day 0 ~ 1 (duration missing in database; assumed 1 day)

culture “adherent” human PSC in “E8”. For ECM, use “Vitronectin” at “N/A” “N/A”.

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

Step 2: mesoderm 1

day 1 ~ 2

culture “adherent” human PSC in “E6 medium” for “1” 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 “1” days.

Step 3: endothelial 1

day 2 ~ 3

culture “adherent” human PSC in “ECGM” for “1” days. For ECM, use “Collagen IV” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “EC” should express “VECad”, “PECAM1”, “VWF”, “KDR”, “NOS3” at the end of culture.

Step 4: endothelial 2

day 3 ~ 5

culture “adherent” unknown species cell in “ECGM” for “2” days. For ECM, use “Collagen IV” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.