The ACVR1 R206H mutation found in fibrodysplasia ossificans progressiva increases human induced pluripotent stem cell-derived endothelial cell formation and collagen production through BMP-mediated SMAD1/5/8 signaling

DOI: 10.1186/s13287-016-0372-6

Authors: Emilie Barruet; Blanca M. Morales; Wint Lwin; Mark P. White; Christina V. Theodoris; Hannah Kim; Ashley Urrutia; Sarah Anne Wong; Deepak Srivastava; Edward C. Hsiao

Step 1: iPSC/ESC maintainance

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

culture “adherent” human PSC in “mTeSR”. “PSC” should be “unspecified regulation” for “ACVR1”. For ECM, use “Matrigel” at “0.15” “g/l”. Medium should contain factors shown in the following table.

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

Step 2: mesoderm 1

day 1 ~ 5

culture “spheroid” human PSC in “StemPro” for “4” days. “PSC” should be “unspecified regulation” for “ACVR1”. Medium should contain factors shown in the following table, and should be changed once in “4” days.

Step 3: mesoderm 2

day 5 ~ 7

culture “adherent” human PSC in “StemPro” for “2” days. “PSC” should be “unspecified regulation” for “ACVR1”. For ECM, use “Collagen” 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 “PECAM1”, “KDR” at the end of culture.

Step 4: endothelial 1

day 7 ~ 10

culture “adherent” human EC in “Endothelial Cell Medium” for “3” days. “EC” should be “unspecified regulation” for “ACVR1”. For ECM, use “FN1” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “3” days.

Ideally, “EC” should express “PECAM1”, “CDH5”, “KDR”, “VWF”, “TEK”, “CD34” at the end of culture.